Welcome to the Science2010 Undergraduate Research Poster

Reception at the University of Pittsburgh

We would like to take this opportunity to thank the undergraduate researchers and their faculty

mentors for sharing their scholarship with the University community at this event.

Undergraduates from the School of Arts and Sciences, the Swanson School of Engineering, the

School of Health and Rehabilitation Sciences, the School of Nursing and the Honors College are

presenting at this reception.

We hope you enjoy the presentations and take the opportunity to engage our undergraduate

researchers in conversation about their work. Inside this program you will find research abstracts

written by the undergraduates.

We wish to thank Science2010 for graciously including this undergraduate research event in its

2010 program.

Office of the Provost

University of Pittsburgh

For more information about Undergraduate Research at Pitt, www.undergradresearch.pitt.edu

Suyesh Acharya

Bioengineering

Swanson School of Engineering

Faculty Mentor: Dr. Kong Chen and Dr. Robert Brychta, NIH

USING ACCELEROMETERS TO DETERMINE HUMAN MOVEMENT

Accurately and non-invasively measuring energy expenditure in the free living environment is a

challenging task, but recent enhancements in size, memory, and performance has allowed

accelerometers to become increasingly used for this purpose. Although the accelerometer method lacks

the ability to precisely differentiate between similar activities like static postures and walking motions,

improvements in detection of these events will increase the accuracy of energy expenditure models, and

may have broader applications in behavioral monitoring. This study was performed to develop a model

to identify the human movements most prominent in free living using raw accelerometer signals. Six

males and nine females, aged 19-50 years, with height 153.5-186.6 cm and weight 54.3-85.6 kg

participated in the following activities in varied order: Walking Upstairs, Walking Downstairs, Level

Walking, Sitting Straight, Sitting Lean, Free Sitting, Laying Flat, Free Laying, and Jogging. Raw data

collected from a tri-axial hip-worn accelerometer (ActiGraph GT3X) was used to develop an algorithm.

Each activity was repeated several times to provide both “training” and “validation” data for the

algorithm. The algorithm easily distinguished between low, moderate, and high intensity activities,

identifying Sitting, Jogging, and Laying with high accuracy (above 86%) and Walking Downstairs, Walking

Upstairs, and Level Walking with moderate accuracy (above70%). The study shows great potentials for

detecting different human locomotion and postures from raw accelerometer signals. Testing even more

subjects of a wider range of age and size at broader and faster walking speeds will allow for a more

concise and accurate energy expenditure model to be developed.

Bahar Ahani

Bioengineering

Swanson School of Engineering

Faculty Mentor: Dr. Mitra Lavasani

ROLE OF MUSCLE-DERIVED STEM CELLS IN A MOUSE MODEL OF AGING

Aged individuals have a dramatically increased risk of numerous debilitating diseases including bone

fractures, sarcopenia, cardiovascular diseases, diabetes and cancer. Growing evidence supports that

DNA damage is one type of cellular damage that promotes aging. Using a mouse model of a human

progeroid syndrome caused by defective DNA repair (ERCC1-deficient mice), we tested our hypothesis

that muscle-derived stem cells (MDSCs) may be affected with aging. A modified preplate technique was

used to isolate MDSCs from Ercc1-/Δ mice (mice expressing 10% of the normal complement of

ERCC1-XPF with a maximum lifespan of 7 months) and Ercc1-/- mice (mice in which ERCC1-XPF is

genetically deleted, with a maximum lifespan of 1 month). Using Live Cell imaging, the proliferation rate

of Ercc1-/- MDSCs proved to be significantly slower when compared to wild-type (WT) MDSCs. WT

MDSCs showed 80% myogenic differentiation while only 46% of Ercc1-/- MDSCs were able to fuse and

differentiate into myotubes in vitro. To further investigate the myogenic potential of MDSCs in vivo, cells

were injected into the gastrocnemius muscle of MDX/SCID mice, a mouse model of Duchenne Muscular

Dystrophy (DMD). WT MDSCs fused with the host muscle fibers to form large dystrophin positive fibers

showing higher regenerative index. Conversely, the Ercc1-/Δ MDSCs were significantly smaller in

diameter and failed to fuse with the host muscle fibers. These results may suggest that the stem cells

inability to proliferate and differentiate may contribute to the dramatically accelerated aging.

Funding: AG-NS-0303-05, NIEHS ES016114, NIH 1R21AG0344907-01A1

Abdul-Kareem Ahmed

Neuroscience

Arts and Sciences

Faculty Mentor: Dr. Bill J. Yates

DISCERNING THE NEURAL CIRCUITRY OF MOTION SICKNESS IN CATS EMPLOYING C-FOS GENE

EXPRESSION

Motion sickness is elicited during conditions where multiple sensory inputs are present that provide

contradictory information regarding body position in space. However, vestibular stimulation alone can

induce motion sickness granted that the sensory inputs differ from those expected. The neural circuitry

that produces motion sickness is not well established. In the present study, we determined the neural

regions where a large fraction of neurons expressed the intermediate early gene c-Fos during motion

sickness elicited by galvanic stimulation of the inner ear. Eight cats were chronically instrumented for

delivery of current to the inner ear on both sides. During the experimental session, 0.5 Hz sinusoidal

stimulation was delivered; the sinusoids on each side were 90 degrees out of phase, to provide a

contradictory input to the two inner ears that is known to result in motion sickness. During the 90-

minute stimulation session, the animals expressed a variety of indicators of motion sickness, including

salivation, retching and immobility. One hour after stimulation was concluded, the animals were

anesthetized and perfused transcardially with paraformaldehyde, and their brains were harvested for

histological analysis. Sections through the brainstem and diencephalon were processed

immunohistochemically to visualize c-Fos. High concentrations of Fos-positive neurons were present in

the nucleus tractus solitarius, respiratory nuclei, periaqueductal gray, vestibular nuclei, parabrachial

nucleus, raphe nuclei, and infratrigeminal nucleus. Presumably, all of these areas participate in

generating motion sickness. Some of these areas contain a high density of serotoninergic neurons; we

confirmed that Fos and serotonin were often co-localized in the same cells.

Funding: NIH, NIDCD

Ogechi Akalegbere

Biology

Arts and Sciences

Faculty Mentor: Deborah Jacobs-Sera

CHARACTERIZATION OF MYCOBACTERIOPHAGE BIGROCH

Bacteriophages were discovered in the early 20th century. With an estimated population of 1031

particles worldwide, these organisms have been used as tools for DNA recombination and genomics as

well as the prevention of bacterial contamination in foods. Understanding the genomic makeup of

bacteriophages and how they interact with their host is important to this investigation.

Mycobacteriophages infect a Mycobacterium host. M. tuberculosis, the causative agent of tuberculosis

(TB), is the prominent pathogen of this genus. Drug resistant strains of M. tuberculosis have become

quite problematic to treat, calling for new approaches. Mycobacteriophages are being investigated as

alternative treatments and as useful tools to manipulate the mycobacterial genomes. Understanding

their host range is important in gathering knowledge on how and why they infect certain bacteria.

Mycobacteriophage BigRoch was isolated from a soil sample collected in 2008. BigRoch was isolated on

the host, M. smegmatis, which is a nonpathogenic relative to M. tuberculosis. At present, BigRoch has

been purified, amplified, and its DNA has been extracted and analyzed. This poster describes

Funding: HHMI

Jordan Anderson

Bioengineering

Swanson School of Engineering

Faculty Mentor: Dr. Bridget M Deasy

STEM CELL MIGRATION ANALYSIS USING ROBOTIC IMAGING SYSTEMS

To study migration processes in vitro, we use a novel time-lapsed microscopy system that allows us to obtain unique data related to cell migration and cell chemoattraction. We used progenitor cells to determine how cells may migrate into a wound, and human mesenchymal stem cells (MSCs) to determine if the system could be used to study chemoattraction. We found that the system could detect significant differences in cell migration, depending on factors including distance from the wound or from the growth factor signal. Migration: The scratch wound assay, in which a confluent layer of cells is wounded by manually scratching a region of the plate to remove all cells, was used. Chemoattraction: Human MSCs were transduced to express a stimulatory growth factor and were cultured and pelleted. Non-transduced cells and the transduced pellet were plated. Live cell time-lapsed imaging was used to capture images at 10-minute intervals(1). Cells on the edge of the wound were observed to infiltrate about 1/3 of the scratch wound area by 36 hours. All cells moved in the direction towards the scratch wound, with no cells migrating inwards toward the confluent region. There was no difference in the mean directional persistence of cell migration for the 2 regions. In the chemoattraction experiment, MSCs in Region 1 appear to display random migration tracks. Some cells in Region 2 appear to exhibit non-random motion in the direction of the chemoattracting pellet. Cell velocity (um /min) was significantly higher for cells nearer to the GF signal as compared to cells farther from the signal. Live cell imaging greatly enhances cell science and tissue engineering by providing a unique dataset to quantify numerous temporal changes in cell populations. Our analysis shows that MSCs chemoattraction to various GFs can be determined readily and quantified to serve as a basis for design of scaffolds which include GF tags.

Yasir Arafat

Chemical Engineering

Swanson School Of Engineering

Faculty Mentor: Edward Lahoda

A COMPREHENSIVE APPROACH TO LIGHT WATER REACTOR FUEL RECYCLING AND LOWERING

RADIOTOXICITY OF HIGH LEVEL WASTE: ASSESSMENT OF TECHNICAL REQUIREMENTS FOR

PARTIOTION TECHNOLOGIES

A proper solution to the growing inventory of spent nuclear fuel has major implications on the future of

nuclear power. The “open cycle” strategy currently adopted in the USA is a threat to the sustainability of

the nuclear option and to its potential growth vs. other energy sources. The current trend can be

reversed by adopting a proper strategy for closing the fuel cycle, with a judicious choice of isotopes to

be separated from used nuclear fuel (UNF) and transmuted by further irradiation cycles. Many specific

solutions have been proposed on this regard. This report is proposing a new approach, which involves

redefining the specifics of the main systems involved (fuel type, reactor design and reprocessing

techniques) to minimize the radiotoxic content of the final high-level waste (HLW) to be disposed. In a

closed fuel cycle, although the transmutation rate of actinides depends on the specific properties of the

system adopted for transmutation (e.g. fast reactors, accelerator-driven systems etc), the final waste

will be predominantly characterized by the reprocessing plants. The efficiency that can be achieved in

the separation and recovery of some critical isotopes is a key to reduce the HLW radiotoxic content. This

paper focuses on identifying these critical isotopes, which are responsible for the long- and

intermediate-term radiotoxicity in the conventional uranium-based fuel cycle and in a proposed

alternative, the thorium-based fuel cycle. The isotopic composition at discharge from fuel typical of

various spectral conditions (thermal and fast range) in either the uranium-based or the thorium-based

fuel cycle has been calculated and its radiotoxic evolution with time analyzed using ORIGEN-ARP. The

analysis reveals that thorium fuel produces negligible amount of transuranics (TRUs), which makes it the

preferred option to minimize the issues associated with their partition and in-reactor transmutation.The

various options available in current state-of-the-art reprocessing techniques and suitability to meet

these specifics will be discussed in the final poster.

Funding: Westinghouse Electric Company

Alison Austin

History and Philosophy of Science

Arts and Sciences

Faculty Mentor: Dr. Nancy Kaufmann

EXPRESSION OF AQP17664 IN LIPID STORAGE SITES SUGGESTS A ROLE IN FAT METABOLISM

Intercellular glycerol transport is crucial for an animal’s ability to maintain homeostasis and is necessary

for survival. However, little is known about possible mechanisms in the cell membrane that transport

glycerol. Evidence suggests that specific types of water channels called aquaporins also allow glycerol

molecules in and out of the cell (aquaglyceroporins). Previous studies on mice involving an

aquaglyceroporin called AQP7 have shown that these channels are involved in mammalian fat

metabolism. AQP7 knockout mice showed impaired ability to release glycerol from adipocytes

(Norikazu Maeda, PNAS). We are interested in learning more about the role of aquaglyceroporins in fat

metabolism during early life stages by using Drosophila melanogaster as a model organism. I have

investigated the expression of an insect homologue of AQP7 called AQP176664 in various drosophila

tissues, focusing on adipocytes found in the fat bodies. Reverse Transcriptase- PCR technique was used

to compare expression of AQP17664 with other aquaporins in the fat bodies of third-instar wandering

larvae (a life cycle stage when the insect is actively feeding and storing fat). Results showed that

AQP17664 is expressed in fat bodies, indicating that it may channel glycerol during fat metabolism. By

learning more about how AQP1766 functions in the transport of glycerol, we can further understand

metabolic pathways of pest insects such as the malaria carrier Anopheles gambia, which relies on stored

glycerol for a source of energy.

Funding: Howard Hughes Medical Institute

Varun Badami

Neuroscience

Arts and Sciences

Faculty Mentor: Dr. Bill Yates

DISTRIBUTION OF HYPOTHALAMIC NEURONS WITH OREXIN OR MELANIN CONCENTRATING

HORMONE IMMUNOREACTIVITY AND MULTISYNAPTIC CONNECTIONS WITH DIAPHRAGM

MOTONEURONS

Prior work showed that neurons in the lateral, dorsal, and perifornical regions of the tuberal and

mammillary levels of the hypothalamus participate in the control of breathing. The same areas also

contain large numbers of neurons that produce either orexins (hypocretins)or melanin concentrating

hormone (MCH). These peptides have been implicated in regulating energy balance and physiological

changes that occur in transitions between sleep and wakefulness, amongst other functions. The goal of

this study was to determine if hypothalamic neurons involved in respiratory control, which were

identified in cats by the retrograde transneuronal transport of rabies virus from the diaphragm, were

immunopositive for either orexin-A or MCH. In animals with limited rabies infection of the

hypothalamus (<10 infected cells/section), where the neurons with the most direct influences on

diaphragm motoneurons were presumably labeled, a large fraction (28–75%) of the infected

hypothalamic neurons contained orexin-A. In the same cases, 6–33% of rabies-infected hypothalamic

cells contained MCH. However, in animals with more extensive infection, where rabies had presumably

passed transneuronally through more synapses, the fraction of infected cells that contained orexin-A

was lower. The findings from these experiments thus support the notion that hypothalamic influences

on breathing are substantially mediated through orexins or MCH.

Funding: NIH grants 5R01-DC003732-11 and 3R01-DC003732-11S1, and P40-RR-018604 from NIH's

National Center for Research Resources

Rebecca Belan

Mechanical Engineering

Swanson School of Engineering

Faculty Mentor: Adrian Michael

ENZYMATIC SENSORS FOR DETECTION OF NEUROTRANSMITTERS IN OXYGEN DEPRIVED TISSUE

Electrochemical detection of neurotransmitters is an important method for understanding the brain.

Single carbon fiber electrodes are widely used as their small size allows for detection without causing

damage to the brain. In conjunction with enzyme trapping redox hydrogels, electrode arrays (daggers)

are especially useful for the detection of neurotransmitters that are not themselves electroactive. To

date no array has been made of similar scale to a carbon fiber electrode. For the past year, the Michael

Group has been developing a triple-band platinum electrode array with aims of eventual in vivo use. To

date, daggers with platinum bands with approximately 10x20&#956;m cross sections have been

produced. Previous work has established a procedure for manufacturing free standing polymer (Su8)

structures through the use of lithography and aluminum as a dissolvable layer between the Si wafer and

Su8 structure (“lift-off”). The structures produced through this procedure have a cross section of the

desired size, and further corrected the curling issue and stiffened the electrodes. Signals have been

successfully read in a potassium ferrocyanide electrochemical reaction and testing is continuing on to

various neurotransmitters. This paper focuses on a redesign of the electrodes, to further stiffen them

and make them easy to handle, as well as on a design of a holder for testing the electrodes.

Funding: NIH #MH075989

Taylor Bissell

Bioengineering

Swanson School of Engineering

Faculty Mentor: Dr. Bridget Deasy

METHODS TO ISOLATE AND IDENTIFY QUIECSENT HUMAN MUSCLE DERIVED STEM CELLS

Stem cells populations are heterogeneous and contain dividing and non-dividing subpopulations.

Included in the non-dividing subpopulation are quiescent cells, which are cells that are not currently

dividing, but do have the potential to reactivate and re-enter the cell cycle. Quiescent cells are believed

to be the most potent cells, and are thought to be the cells that survive transplantation during

regenerative medicine, and contribute the most during the regeneration process. Human muscle

derived stem cells (hMDSCs) were labeled with 5-chloromethylfluorescein diacetate (CMFDA) and sorted

using flow cytometry into CMFDA positive (non-dividing) and negative (dividing) populations. The cells

were observed using live cell imaging for 6 days, and the sequential jpegs were analyzed for proliferation

data using image analysis software. Cells were also followed to determine lineage history and to

determine if a cells was actively dividing. The sorted cells were cultured for a long-term expansion to

determine longevity. The proliferation data used to determine the fraction of dividing cells in each

subpopulation. 73% of the dividing cells were actively dividing, while 51% of the non-dividing cells were

actively dividing. After 40 days in culture, both subpopulations were expanding at a similar rate with

12.4 doublings each. Identification of the subpopulations allows for an in depth understanding of the

subpopulations. The quiescent cells may have the greatest proliferative capacity due to the fact that

they are not actively dividing, and after transplantation, give rise to reparative progeny for successful

regenerative medicine therapies.

Funding: National Institute of Arthritis and Musculoskeletal and Skin Disease (R03 AR053678)

Jennifer Boles

Neuroscience

Arts and Sciences

Faculty Mentor: Amy K. Wagner, MD

LEPTIN ASSOCIATIONS WITH POST TRAUMATIC SEIZURE AFTER SEVERE TRAUMATIC BRAIN INJURY

Posttraumatic seizures (PTS) are a common consequence of traumatic brain injury (TBI) and are also

associated with mortality and poor outcome after injury. Leptin, a 16-kDa adipose derived hormone is

known for its effects on metabolism, energy balance, reproductive regulation, the immune response,

and neuroprotection. It has also recently been found to play a role in seizure activity. Rat models have

been used to show that leptin prevents seizures by inhibiting hippocampal neurons by activating a large

amount of Ca2+-activated K+ (BK) channels. However, in proconvulsant rodent models, leptin enhances

NMDA receptor-mediated increases in intracellular Ca2+ levels and synaptic transmission in rat

hippocampal cell cultures and brain slices. The purpose of this study was to explore the associations

between leptin levels and the occurrence of PTS after TBI. This retrospective study included 145 adult

Caucasians with a severe TBI. Time to first seizures and BMI were determined through a medical chart

review. Leptin levels were determined with Cerebrospinal fluid (CSF) which was collected twice a day

for 5 days post injury. Approximately 25% of the population experienced at least one documented

seizure after injury. Higher mean leptin levels were associated with PTS occurring within one month

post-TBI (p=.005). In addition, the average BMI for those who seized within a month was significantly

higher than the average BMI for those who did not seize within the time period (p=.046). It appears that

elevated average weekly leptin levels are associated with an increased risk of developing PTS.

Funding: DODW81XWH-071-0701

Sameul Brayer

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Amy K. Wagner

EXAMINING LEARNING STRATEGIES WITH THE MORRIS WATER MAZE SPATIAL MEMORY PARADIGM:

A COMPARATIVE ASSESSMENT IN EXPERIMENTAL TRAUMATIC BRAIN INJURY

Historically the Morris water maze (MWM) has been utilized as a tool for assessing spatial learning and

memory in rodents, by allowing animals to swim to an escape using proximal and distal cues for

navigation. Questions remain about whether animals rely entirely on external cues for navigation, a

spatial strategy, or also incorporate non-spatial search strategies, the effects of which may influence

overall performance. We hypothesized that non-spatial pre-training prior to MWM testing would result

in lower latencies for both acquisition and reversal trials during spatial testing. We further hypothesized

that pre-training does not need to occur immediately prior to spatial testing and that training prior to

the controlled cortical impact (CCI) model of traumatic brain injury (TBI) can be beneficial to MWM

spatial memory performance two weeks post-injury. 39 Sprague-Dawley rats underwent CCI

(2.8mm;4m/s) or sham surgery. 21 rats received non-spatial pre-training that was performed for 4 days

prior to surgery with a dynamic hidden platform and static entry point. Motor testing occurred on post-

surgery d1-d6 to ensure that motor ability returned to pre-surgical levels. MWM spatial acquisition trials

were performed beginning 14d post-surgery. Significant improvements in swim latencies for acquisition

trials were observed for the pre-trained CCI group compared to CCI rats without pre-training. This

suggests that pre-training facilitated the use of learned behavioral strategy that was able to be utilized

in rats with CCI. The work also highlights the importance of extra-hippocampal training and function as

a relevant compensatory strategy for spatial learning in experimental TBI.

Funding: Department of Physical Medicine and Rehabilitation, University of Pittsburgh

Christopher Brett

Neuroscience

Arts and Sciences

Faculty Mentor: Amy K. Wagner, MD

OUTCOME AND INJURY SEVERITY ANALYSIS OF QUANTITATIVELY CATEGORIZED HYPOGONADOTROPIC

HYPOGONADISM IN A POPULATION IN SEVERE MALE TRAUMATIC BRAIN INJURY (TBI) PATIENTS

Research has demonstrated derangements in chronic gonadal/adrenal serum hormone profiles in the

first week following a TBI. Testosterone (TEST) and leutinizing hormone (LH) decline acutely to chronic

depletion suggesting hypogonadotrpic hypogonadism. The purpose of this study was to examine

hypogonadotropic hypogonadism serum hormone profiles in a male population after TBI with respect to

outcome and injury severity. Male TBI subjects (N=38) had average hormone levels from blood samples

taken acutely, every two weeks for the first 6 months and after 12 months post-injury This study defined

hypogonadotropic hypogonadism quantitatively based on depleted TEST and LH serum levels with

respect to normal hormone ranges according to the UPMC Laboratory. Injury severity and outcome

were compared between hypogonadotropic hypogonadic subjects (HHS) and non-hypogonadotropic

hypogonadic subjects (NHS). Injury severity measures included the Glasgow Coma Scale (GCS) and Injury

Severity Scale (ISS). Outcome measures included Glasgow Outcome Scores (GOS), Disability Rating Scale

(DRS), Functional Independence Measure (FIM), PHQ-9 depression scale, Quality of Life (QOL), and a

cognitive composite measure. Injury severity was not significantly related to hypogonadism. Six month

GOS, DRS, FIM, and cognitive composite scores for HHS were significantly worse (P<0.05) than NHS.

Depression and QOL were not significantly different between HHS and NHS. NHS TEST, LH and E2 levels

were consistently higher than HHS in the chronic phase. Male TBI patients with hypogonadotropic

hypogonadism have worse outcomes and abnormal hormone levels chronically after injury.

Funding: Centers for Disease Control and Prevention (#R49/CCR323155), Department of Defense

(#W81XWH-07-0701), & National Institute of Health (#P50NS030318)

Stephen Canton

Bioengineering

Swanson School of Engineering

Faculty Mentor: Debora Jacobs-Sera

ISOLATION, PURIFICATION, AND CHARACTERIZATION OF MYCOBACTERIOPHAGE AKOMA

Bacteriophages are viruses that infect bacteria. With over 1031 particles in existence, bacteriophages

represent the most prevalent pool of unknown genetic information. These organisms are commonly

found in areas with large populations of bacterial hosts. Mycobacteriophage Akoma, was isolated from

a soil sample the North Oakland Area of the University of Pittsburgh campus. Using Mycobacterium

smegmatis mc2155 as a host, my project is the characterization of Mycobacteriophage Akoma. This

novel bacteriophage has been purified and prepared for sequencing. Once analyzed, its genome will be

compared to over one hundred mycobacteriophages. The plaque morphology, physical structure, and

restriction enzyme digest suggest that Akoma belongs to Cluster B. Using cluster specific primers,

Akoma is believed to be in subcluster B3. This poster describes the physical and molecular

characteristics known to date about Mycobacteriophage Akoma.

Funding: Howard Hughes Medical Institute

Aaron S Cantor

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Susan Kalisz

GARLIC MUSTARD-INVADED FOREST SOILS SUPPRESS FUNGAL HYPHAL GROWTH

Garlic mustard (Alliaria petiolata, Brassicaceae), hereafter GM, is a widespread invader of North

American forests. Like some members of the mustard family, GM increases soil levels of allyl

isothiocyanate (AITC), a powerful anti-fungal agent suspected to increase GM’s invasive success. Low

AITC concentrations are known to inhibit fungal spore germination in vitro, but the effect of GM’s AITC

on fungal growth within invaded soils is unknown. Therefore, we buried cellulose membranes in field

plots with no GM (control) or with natural densities of GM at three sites for four weeks and then

quantified hyphal colonization on the membranes using digital image analysis. We found that total

hyphal length is significantly lower in GM vs. control plots, and mean hyphal length/image is significantly

lower in GM vs. control plots at two sites. These results show that GM-infested soils suppress vital

fungal growth and implicate that AITC produces this effect.

Funding: NASA Pennsylvania Space Grant Consortium

Evan Carder

Chemistry

Arts and Science

Faculty Mentor: Dr. Kazunori Koide

STUDIES TOWARD AN IDEAL FLUORESCENCE METHOD TO MEASURE PALLADIUM IN FUNCTIONALIZED

ORGANIC MOLECULES

Detection and removal of toxic metals are crucial steps in the preparation of organic compounds. These

steps, involving active pharmaceutical ingredients (APIs) and bioactive compounds, are particularly

important for safe pharmaceutical practice and reproducible biological studies. However, detection of

metals in synthetic organic compounds cannot always be achieved by currently used techniques, such as

inductively coupled plasma mass spectroscopy (ICP-MS). False negative data for toxic metals are of

particular concern due to the dangerous possibility of exposing the general public to tainted

pharmaceuticals. Therefore, in order to ensure public safety, an alternative approach for quality control

of synthetic compounds is warranted. Palladium, a common metal used in numerous organic reactions,

is frequently found in synthetic compounds due to poor removal using typical purification protocols.

Because ICP-MS is not likely to be a unified solution for the detection of palladium in all synthetic

compounds, it is important to develop alternative methods that are sensitive, specific, rapid, and

inexpensive. Furthermore, such methods must be able to detect residual palladium in the presence of

APIs in a 5-10 ppm range, which is suitable for government regulations. Herein, we report an ideal

fluorescence detection method, based off of the Tsuji-Trost deallylation reaction, which meets the

necessary requirements for the detection of palladium in functionalized organic molecules. In our study,

we account for the effects of sodium borohydride, temperature, phosphine ligand, and phosphate ions

in kinetics. Such study has applications for synthetic chemists, pharmacists, and patients who are

subjected to palladium-based drugs.

Funding: NSF

Anne Caruso

Neuroscience

Arts and Sciences

Faculty Mentor: Monica A. Perez

THE EFFECT OF FORCE DIRECTION ON MOTOR SKILL LEARNING

In healthy humans, bilateral isometric index finger forces in opposing directions exert differential

changes in motor cortical function (Yedimenko and Perez 2010). It remains unknown whether the

direction of force in one hand could influence motor skill learning by the contralateral hand. In this

study, we examined the effect of index finger isometric forces in two different directions (abduction and

adduction) with the right hand on the magnitude of motor skill learning by the left hand. During training,

subjects were instructed to move a cursor following a series of target lines on a computer screen by

performing 0-10% of their maximal abduction force with the left index finger while the right hand

completed 30% of isometric index finger abduction or adduction. Each training period consisted of 10

sessions of practice. Within each session, three different targets were randomly presented eight times

with a total of 24 figure presentations. There were 2 minutes of rest after each session was completed.

We found that motor performance in the left hand improved to a larger extent when the right hand was

performing abduction compared to adduction force. Similarly, the amount of error measured in the left

index finger was significantly decreased when the right index finger completed abduction compared to

adduction force. Our results suggest that motor skill learning in the non-dominant hand can be

differentially affected by the direction of the force exerted by the contralateral hand.

Funding: National Institutes of Health

Brittany Charsar

Biological Sciences

Arts and Sciences

Faculty Mentor: Joe Martens

ISOLATION AND CHARACTERIZATION OF SPT16 MUTANTS THAT DEREPRESS SER3

Intergenic DNA comprises a major portion of eukaryotic genomes and is highly transcribed. Recently,

transcription of intergenic DNA, leading to a non-coding RNA (ncRNA) product, has been shown to be a

major player in the regulation of coding genes. Our lab has identified one ncRNA, SRG1, which has a

regulatory role in gene expression, where the act of transcribing SRG1 represses its adjacent gene, SER3.

In addition to SRG1, the essential yeast protein Spt16 plays a part in this repression of SER3 by

maintaining nucleosome structure. Spt16 joins with other members of the FACT complex to reposition

nucleosomes around RNA polymerase II during active transcription. Maintaining nucleosome

architecture is important in denying other transcription factors access to SER3 promoter elements. To

investigate the mechanism by which Spt16 represses SER3, a genetic screen was performed to isolate

mutations in SPT16 that derepress SER3. spt16 mutants, generated through PCR-based mutagenesis, are

being subcloned and retested for derepression of SER3 using a SER3-HIS3 reporter construct. Mutants

that pass re-screening are then subjected to Northern analysis to assay endogenous levels of SER3. To

begin categorizing spt16 mutants that derepress SER3, we are measuring Spt16 protein levels by

Western analysis and testing for other phenotypes by performing growth assays on a variety of different

media. Thus far, seven mutants have been identified that are expressed at wild-type levels and are

specific to the regulation of SER3. Future experiments to characterize these Spt16 mutants will help us

gain insight into how Spt16 functions mechanistically.

Funding: NIH

Olivia Creasey

Bioengineering

Swanson School of Engineering

Faculty Mentor: Dr. Rocky Tuan

EFFECT OF PARTIAL REPROGRAMMING ON IMMUNOPHENOTYPE OF ADULT STEM CELLS

Many tissue engineering approaches use adult mesenchymal stem cells (MSCs) to reconstitute or repair

damaged tissues. For cartilage tissue engineering, a large number of stem cells are required. Lengthy in

vitro expansion results in phenotypic changes in this cell population, namely decreased proliferation,

down-regulation of MSC-associated surface markers, and decreased differentiation potential. We

investigated how in vitro expansion changes the immunophenotype of MSCs, and whether partial

reprogramming, which is the process of moving an adult cell along the pathway from limited

multipotency toward pluripotency without returning it to a completely pluripotent state, returned late-

passage cells to their early passage immunophenotype. MSCs from hip arthroplasy specimens with a

passage number P8 or greater (late-passage MSCs) or P3 or less (early-passage MSCs) were fixed,

permeabilized, and immunostained with antibodies to CD34, CD44, CD45, CD73, CD90, CD105, CD146,

Oct4, Sox2, Nanog, SSEA3, SSEA4, TRA-1-60, TRA-1-81 (BD), Stro1 (BioLegend), and c-Myc (Abcam) to

generate an immunophenotype. Immunostained cells were analyzed on a FacsDiva (BD), and the marker

positivity was determined by analysis in FlowJo. Significant differences were determined using a two-

tailed student’s t-test. Nonstatistically significant trends were observed suggesting differences between

early-passage and late-passage cells for the markers CD105 and CD146, and in particular SSEA4, which

was higher in late-passage cells (96.5% positive) than in early-passage cells (3.04% positive). We

observed no significant differences as a result of partial reprogramming for the markers that we

examined. With a broader panel of markers, this technique may prove useful for selecting partially

reprogrammed cells.

Fuding: Commonwealth of Pennsylvania Department of Health

Christopher Davis

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Karen Arndt

GENETIC ANALYSIS OF THE HISTONE MODIFICATION DOMAIN OF THE PAF1 COMPLEX SUBUNIT RTF1

Transcriptional regulators have been shown to cause alterations in chromatin structure through various

modifications to the nucleosomal histone proteins, such as acetylation, methylation, and ubiquitylation.

The Paf1 complex is a group of transcription elongation factors, consisting of Rtf1, Cdc73, Ctr9, Leo1,

and Paf1, that associates with actively transcribing RNA polymerase II. Rtf1 is required for

monoubiquitylation of lysine 123 in histone H2B, which subsequently directs the methylation of lysine

residues in histone H3. Rtf1-dependent histone methylation aids in the maintenance of proper

chromatin structure during transcription elongation. We have investigated the role of Rtf1 in this

process by observing the effects of several different amino acid substitutions made within Rtf1.

Particularly, these substitutions lie within a region known as the histone modification domain (HMD).

The protein sequence of the HMD is highly conserved from the unicellular eukaryote S. cerevisiae to

more complex organisms, including humans, suggesting its importance in eukaryotic transcriptional

regulation. Accordingly, amino acid substitutions in the HMD lead to phenotypes associated with

defects in transcription, including 6-azauracil sensitivity, loss of telomeric silencing, derepression of

cryptic initiation, and an Spt- phenotype. We have assessed the ability of particular substitutions to

disrupt methylation of histone H3 as well as ubiquitylation of histone H2B. We also show that the HMD

is sufficient for ubiquitylation of histone H2B. Current work involves investigation of possible sources of

extragenic suppression of rtf1 mutant phenotypes identified through a mutagenic suppressor screen.

Funding: National Institutes of Health and the Arnold and Mabel Beckman Foundation

Kyle DeHart

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Tia-Lynn Ashman

THE GREAT STRAWBERRY CAPER: SCIENTIFIC CURRICULUM DEVELOPMENT

Understanding science requires hands-on learning. We aimed to initiate the development of a biology

curriculum that local area middle school teachers can utilize in their classrooms to teach students

genetics and ecology. The basic premise is to use strawberries, purchased from local food markets, as a

model system for hands-on, lab oriented activities. We began by addressing two questions:1. Is there

phenotypic variation in store-bought strawberries? And is it attributable to the companies that supply

them? If so it might indicate that they are different cultivars. 2: Can we use diagnostic microsatellite

markers to a) successfully differentiate between berries from different companies if they indeed differ,

or b) identify the cultivars? To answer these questions data was collected for several morphometric

characters of berries and DNA was extracted from their sepals. PCR was conducted using two

microsatellite primers known to be useful for fingerprinting strawberry cultivars and results were

analyzed using GeneMapper. We discovered that there was a wide range of variation in strawberry size,

seed number and dimensions, but that only the size differed significantly between the company sources.

The results also showed that quality DNA could be extracted from the sepal tissue found on store-

bought strawberries, even if it was old. Both microsatellite markers amplified products in the expected

range in some of the samples. Combined these results provide promising framework for continued

development and implementation of the middle school curriculum.

Fudning: Howard Hughes Medical Institute

Carly Deibler

Industrial Engineering

Swanson School of Engineering

Faculty Mentor: Dr. Mary Besterfield-Sacre

EFFECTS OF TEAM DYNAMICS BEHAVIORS ON DESIGN ARTIFACT SCORES

The process of creating an innovative product for the first time in a design class can be a challenge for

any student. While knowledge and diligent work are necessities, successful creation of the innovative

product is also a function of team interactions. I examine which components of team interactions affect

design artifact scores. I assert that positive team dynamics during different phases of the design process

leads to higher-quality design and innovative projects. My objective was to identify the effects of three

team dynamics behaviors on various design artifact scores. I also wanted to identify which

subcomponents of team dynamics are most important to a design and at which stages of the project. I

categorized team comments into the areas of team dynamics related to communication, collaboration,

and decision making. I compared their team dynamics information to the actual scores received on their

final product design to determine what, if any, relationship exists between the team interactions and

the artifact scores. Data from 26 senior design teams from the University of Pittsburgh and the Rose-

Hulman Institute of Technology was used in this analysis. My analysis also included a look at which

phases of the project were crucial to team interaction in relation to the final output scores. The output

scores on the final projects included the following criteria: Technical Performance & Standards,

Documentation, Innovation, Working Prototype, and Overall Impact. I also examined the sum of these

scores and the size of the teams in relation to the team dynamics categories.

Funding: Undergraduate Research Internship

Raphael Del Giorno

Neuroscience

Arts and Sciences

Faculty Mentor: Dr. Ariadne Letra

RECENTLY DESCRIBED MATRIX METALLOPROTEINASES ARE EXPRESSED DURING MOUSE

PALATOGENESIS

Matrix metalloproteinases (MMPs) are a family of proteolytic enzymes responsible for degradation of

the extracellular matrix and tissue remodeling during including embryonic development and

palatogenesis. There are 23 MMPs described in humans so far which have been classified into different

groups based on a combination of amino acid sequence homology, presence of specific peptide

structural domains, and in vitro substrate specificity. MMPs are usually divided into collagenases,

gelatinases, stromelysins, matrilysins, membrane-type metalloproteinases, and others, which include

the more recently identified MMPs. Many MMPs have been detected during murine palatogenesis

although no information is available about the expression of the more recently identified MMPs. We

verified gene and protein expression of MMPs not yet described in mouse craniofacial tissues through

real-time PCR and immunohistochemistry (IHC). For real-time PCR, we used full-length first strand cDNA

derived from head and palate of mouse embryos at embryonic days (ED) 12, 13, 14, 14.5, and 15 and

Sybr Green chemistry. Beta-actin was used as an endogenous control. Primers were designed using

Primer3 software. For IHC, we used mouse head tissue sections at ED 13-15. The results showed

differential temporal and spatial expression of MMPs 7, 10, 16, 25, 27, and 28 at critical stages for

palatal development in mice, and correlate with the distribution of their substrates. Hence, the MMPs

investigated in this study might also play a role during murine palatogenesis.

Funding: NIH/NIDCR R00-DE018954

Cassandra Delp

Acute & Tertiary Care

School of Nursing

Faculty Mentor: Dr. Mary Beth Happ

EXPLORING FAMILY’S PERCEPTIONS OF PATIENT COMMUNICATION IN THE ICU

Development and testing of effective communication strategies for critically ill patients who are unable

to speak demands a better understanding of family perceptions of communication. To describe

perceptions of communication by visiting family and friends regarding patient communication in the

ICU. DESIGN: Cross-sectional self-administered survey of 133 family members or friends (visitors) of

awake nonspeaking adult patients (orally intubated or tracheostomy without speaking valve). Family

perceptions of communication quality, satisfaction with communication, and common methods used to

communicate with their family member using a nine-item five point Likert type survey, administered

&#8805; 48 hours after patient admission to ICU. We received 57 completed surveys (response rate

=42.9%). Three-quarters of respondents were women (n= 43), and were related to the patient as spouse

(n= 20; 35%), sibling (n=16; 28%), and adult child (n=16, 28.1%). Respondents were equally satisfied

(n=25) and dissatisfied (n=25) with communication. Forty nine percent (n=28) reported feeling

uncomfortable communicating with the patient. Most reported feeling frustrated (n=36, 62%) and

helpless (n=37, 64.0%) by not being able to communicate effectively with the patient. Although

satisfaction ratings are equivocal, families report discomfort with communication and feelings of

frustration and helplessness with the patient’s impaired communication. The proportion of poor

satisfaction ratings illustrates the need to develop and test new approaches for communication in this

setting to reduce these negative experiences.

Funding: Robert Wood Johnson Foundation Interdisciplinary Nursing Quality Research Initiative grant #

#66633 (Happ MB & Barnato AE)

Vincent DeStefino

Neuroscience

Arts and Sciences

Faculty Mentor: Dr. Bill Yates

RESPONSES OF NEURONS IN THE ROSTRAL VENTROLATERAL MEDULLA DURING CHANGES IN

POSTURE: COMPARISONS BETWEEN CONSCIOUS AND DECEREBRATE ANIMALS

A region of the brainstem, the rostral ventrolateral medulla (RVLM), controls sympathetic nervous

system activity and blood pressure. Prior studies in decerebrate or anesthetized animals showed that

the RVLM integrates a variety of inputs, including those from baroreceptor and vestibular afferents; the

latter signals are presumably involved in regulating blood distribution in the body during postural

alterations. However, no prior studies have considered activity of RVLM neurons in conscious animals.

The goal of the present study was to compare responses of RVLM neurons to moderate-amplitude body

tilts in conscious and decerebrate cats. Single unit recordings were performed on RVLM neurons in 3

awake and 8 decerebrate cats. Individual neurons were assayed for 1) baroreceptor inputs and 2)

modulation of activity during natural vestibular stimulation. A large fraction of RVLM neurons (51/110)

responded to <15° tilts in decerebrate cats. In contrast, <2% of RVLM neurons in conscious animals

responded to the same stimuli. Responses to vestibular stimulation were similar for cells that received

baroreceptor inputs (exhibited cardiac-related activity or responded to carotid artery stretch) as those

without such inputs. These data suggest that responses of RVLM neurons to body tilts are masked in

conscious animals, perhaps because they are not beneficial during moderate-amplitude tilts. Future

studies will examine the mechanism responsible for this response gating, and whether it can be

modified during situations when postural changes that compromise blood pressure are more likely.

Fudning: National Institutes of Health (NIH)

Shannon Donofry

Psychology

Arts and Sciences

Faculty Mentor: Dr. Kathryn Roecklein

ASSOCIATIONS OF DRD4 7R AND DRD2 TAQ1A POLYMORPHISMS WITH BINGE EATING IN PATIENTS

WITH SEASONAL AFFECTIVE DISORDER

The present study aims to investigate the relationship between variations in dopamine related genes,

binge eating behavior, and weight gain in patients with seasonal affective disorder (SAD). Researchers

have proposed the Reward Deficiency hypothesis to explain dopamine function as it relates to addictive

behaviors such as overeating. According to this model, individuals whose dopaminergic systems are

downregulated engage in rewarding behaviors to compensate for this deficit. To extend the work of

Levitan (2004a), the Taq1A A1 restriction fragment length polymorphism (RFLP) linked to the dopamine-

2 receptor gene has been included in the assay, in addition to the 7-repeat allele (7R) of the dopamine-4

receptor gene (DRD4), as both of these polymorphisms have been associated with blunted dopamine

response and higher maximal lifetime body mass index (BMI) in multiple samples (i.e. Chang et al.,1996;

Kaplan et al., 2008; Noble, 2000; Stice et al., 2008; Wang et al., 2001).Sixty- four male and female

participants completed the SIGH-SAD, a modified version of the Hamilton Depression Rating Scale that

includes a subscale structured to ascertain the presence of atypical depression symptoms seen in SAD.

Additionally, participants completed the Questionnaire on Weight and Eating Patterns (QWEP), a self-

administered survey designed to assess binge-eating behavior as defined in the DSM-IV (APA, 2000).

Participants’ height and weight were also measured in order to calculate BMI. DNA was collected using

Oragene DNA self-collection units, and amplified with polymerase chain reactions (PCR). DRD4 PCR

products were visualized through gel electrophoresis, while Taq1A genotypes were determined by

utilizing high-resolution melt analysis (HRMA). Analysis and data collection is ongoing.

Tawny Duliba

Psychology and Neuroscience

Arts and Sciences

Faculty Mentor: Dr. Joseph H. Ricker

EFFECT OF PLEASANTNESS IN EPISODIC MEMORY WITH TBI AND HEALTHY POPULATIONS

Pleasant events have been found to be recalled better than unpleasant events by undergraduate

volunteers (Walker, Vogl, and Thompson, 1997). The research has involved intact, healthy individuals,

however it is not known whether individuals with Traumatic Brain Injury (TBI) exhibit the same pattern.

This population supports this case because individuals with TBI are known to have memory complaints.

Fifteen individuals with TBI (10 males) and thirteen intact controls (11 males) with no significant

differences of age or education participated in a functional Magnetic Resonance Imaging (fMRI) study of

episodic memory. They were presented multiple visual stimuli, such as words, shapes, letters, and

pictures, and asked to rate the items as pleasant or unpleasant. The two populations were presented

the stimuli again and were asked if they were reoccurring or not. Response times and accuracy values

were collected during the recognition blocks and were then sorted based on whether the stimuli were

rated as pleasant or unpleasant. In the TBI and healthy populations there was no significant difference

of response time or accuracy between the groups. However, there was significance with pleasant

responses being more accurately recalled. This study reinforces the idea that pleasant stimuli are more

likely to be recalled than unpleasant stimuli and shows that this pattern can be found in persons with

TBI.

Funding: NIH #R01 NS048178-01

Karthik Dwarki

Chemistry

Arts and Sciences

Faculty Mentor: Drs.Tony M. Plant and Paula Grabowski

HYPOTHALAMIC NEURONS EXPRESSING KISSPEPTIN, NEUROKININ B AND DYNORPHYN (KNDY

NEURONS) CONTROL THE TIMING OF PUBERTY IN THE RHESUS MONKEY, A REPRESENTATIVE OF

HIGHER PRIMATES.

Gonadotropin-releasing hormone (GnRH), a hypothalamic peptide that is secreted in a pulsatile fashion,

provides the drive for the pituitary-gonadal axis that initiates puberty. Interestingly, in higher primates,

robust GnRH pulsatility is also found in infancy. Thus, two hypothalamic postnatal switches control the

timing of puberty in these species: one to turn off GnRH during infancy and the second to reactivate

pulsatility late in juvenile development. In man, mutations of GPR54 are associated with delayed or

absent puberty. The ligand for GPR54 is kisspeptin (KP), which is encoded by KISS1. In monkeys,

expression of KISS1and release of KP in the hypothalamus increase at puberty. KP neurons also express

neurokinin B (NKB), and mutations in this ligand and its receptor in man result in a phenotype similar to

that seen in GPR54 mutations. The purpose of this experiment was to begin to determine whether

decreased expression of hypothalamic KP and NKB underlies the arrest of GnRH pulsatility in infancy.

Serial coronal 25-&#61549;m hypothalamic sections (paraformaldehyde fixed) of three infant (GnRH on)

and three juvenile (GnRH off) male monkeys were prepared. Sections (250-&#61549;m between each)

were stained for KP and NKB using a cocktail of primary antibodies (sheep anti-kisspeptin at 1:120K and

rabbit anti-NKB at 1:6K) and the neuropeptides were detected with fluorescently tagged secondary

antibodies. KP and NKB immunopositive perikarya were counted using a fluorescence microscope with

a mobile stage allowing for systematic scanning of the arcuate nucleus. In infants, the number of KP

perikarya/section ranged from 50-250, while only 15-150 perikarya/section were seen in juveniles. NKB

was coexpressed in about 20% of the KP positive neurons and demonstrated no developmental change.

These findings indicate that arrest of GnRH pulsatility during infancy, which leads to quiescence of the

pituitary-gonadal axis during childhood, maybe due to a decrease in KP expression in KNDy neurons.

Funding: NIH R01 HD013254 and U54HD08160

Eussera El-Magbri

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Nancy Kaufmann

THE ELUSIVE ROLE OF THE BIG BRAIN AQUAPORIN IN BRAIN FUNCTION OF ADULT DROSOPHILA

MELANOGASTER

Aquaporin-4(AQP-4) is a major cranial water channel near the interfaces of the blood-brain barrier(BBB)

and the brain-Cerebrospinal fluid barrier(CSF) and is linked to the formation of brain edema. A promising

new therapy for treating brain edema is by inhibiting AQP-4 expression. It is reported that Drosophila

melanogaster has an aquaporin named big brain, which has regions of high-sequence homology to AQP-

4. This aquaporin is shown to have a crucial function in fruit fly embryos during neurogenesis by

controlling the number of neural precursor cells. However, big brain does not seem to be a water-

permeable aquaporin. Better yet, little is known about big brain expression in adult flies, a key stage we

chose to study in order to better understand aquaporins in mammalian brain physiology. We carried out

controlled RT-PCR experiments with big brain primers to look for big brain expression in whole male

adult flies, fly heads, and fly brains, and big brain seemed to be found in all three areas, especially the

latter. Yet, the question remains as to why big brain is present in adult Drosophila. With further

experiments, these new findings can potentially help answer the following questions: (1) what is the

elusive function of the big brain “non-water-specific” aquaporin in adult flies, (2) what is the

physiological significance of aquaporins, and (3) how can we control the regulation of fluid transport

through manipulations of AQP-4 expression.

Funding: Howard Hughes Medical Institute

Tracy Fan

Health and Community Systems

School of Nursing

Faculty Mentor: Ann M. Mitchell, PhD, RN, FAAN

ASSESSING A WELLNESS EDUCATION GROUP INTERVENTION (WEGI) FOR ADULTS WITH CHRONIC

SEVERE MENTAL ILLNESS (CSMI)

Chronic severe mental illness (CSMI) is a common disorder in the United States, with adverse effects

that include: poorer physical and mental health, significantly shortened life-expectancy, increased

poverty, and poorer health literacy. Health promotion educational interventions have been shown to be

highly effective, and nursing students are well-qualified to implement a unique Wellness Education

Group Intervention (WEGI). The purpose of this project was to implement and evaluate an 8-week WEGI

in order to educate adults who have CSMI with knowledge related to specific health promotion activities

and skills for disease prevention, utilizing upper-class undergraduate nursing students enrolled in the

psychiatric nursing course. Quantitative data on consumer satisfaction show that the vast majority (94%)

of participants liked something about the program and were glad they came. Qualitative data from

Focus Group review sessions following the WEGI show that participants expressed general satisfaction

with having attended the program. They expressed the desire for the program to continue, while some

participants also expressed the achievement of specific personal health benefits, including healthy

lifestyle modifications. There were three primary areas of self-reported interest and benefit to the

mental health consumers who participated in the WEGI: socialization and support; education and

information; and motivation to change. The WEGI is a successful and easily-implemented wellness

education intervention program for adults with CSMI living in the community. The WEGI could serve as a

model for other multidisciplinary undergraduate training programs and for wellness education in similar

communities.

Jeffrey Fein

Physics and Astronomy

Arts and Sciences

Faculty Mentor: Dr. Steve Dytman

Light Injection Calibration of the Photo-Multiplier Tubes for the MINERvA Detector

MINERvA (Main INjector ExpeRiment for v-A) at Fermilab is an experiment designed to measure

properties of neutrinos, a type of subatomic particle, with atomic nuclei. This is done by shooting a

beam of neutrinos at a detector, where they interact with nuclei. The products of these interactions

deposit energy in the detector, which correspondingly produces light that is converted into an amplified

measurable electrical signal via Photomultiplier tubes (PMTs). The PMT gain, which is the amount of

charge produced per input photon of light, must be measured to calibrate the detector. To measure the

gain, MINERvA needs calibration to match electrical signals from 30,000 PMTs to energy deposited by

interaction products. An external Light-Injection system sends controlled pulses of light down fiber-optic

cables into a box where the pulses are diffused over a collection of PMTs. Over the past fifteen months I

worked with the University of Pittsburgh MINERvA group, measuring systematic errors of the gain.

Calibration data was taken from the detector PMTs in compiled intervals of 3 to 15 minutes each for

several months. The comparison of data from consecutive intervals established the gain stability over

short intervals. I then measured the gain stability over longer time intervals and found that it did not

vary significantly from that over short intervals. I determined that the gain was stable over a period of

several weeks.

Funding: Department of Energy

Brandon Fields

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Lewis Jacobson

DISRUPTION OF A MUSCLE ADHESION COMPLEX CAUSES MUSCLE PROTEIN DEGRADATION IN C.

ELEGANS

About one hundred “muscle” genes of C. elegans showed decreased expression levels after prolonged

spaceflight. One of these genes (unc-112) encodes a member of an integrin-containing, transmembrane

attachment complex that forms muscle-muscle attachments, anchors muscle contractile fibers to the

hypodermis, and is highly homologous to human focal adhesion complexes. Knockdown via mutation

(lowers protein function) or RNA interference (lowers amount of protein) of any one of eleven genes

encoding members of this complex provokes protein degradation in muscle cytosol. This result was

consistent for genes whose products reside inside the muscle cell, in the extracellular matrix, or traverse

the muscle PM. Inhibition of either the proteasome or autophagy by drugs or RNAi knockdown failed to

prevent protein degradation in attachment-disrupted mutants, suggesting that a novel protease or a

combination of proteases is responsible. Partial inhibition of degradation resulted when amounts or

functions of other cellular proteases such as calpains, cathepsins, or caspases were decreased

individually or in combination, again suggesting the involvement of multiple proteases. In addition to

muscle protein degradation, which begins about 18-24 hours after acute disruption of the attachment

complex, paralysis occurs in an unc-112ts mutant within 24 hours of disruption. This paralysis can be

prevented by mutation or RNAi knockdown of the immunoglobin-like protein(s) encoded by the gene

dim-1. Our data suggest that either the attachment complexes are highly dynamic structures, or new

attachment complexes must be continuously formed to support body growth even after adulthood is

attained.

Funded by NIH grant 5R01-AR054342. EAO supported by the Pennsylvania Space Grant Consortium, the

Univ. of Pittsburgh, the Biochemical Society (UK), a Barry M. Goldwater Scholarship, and the Howard

Hughes Medical Institute. BDF supported by the Howard H

Heather Friedberg

Computer Science

Arts and Sciences

Faculty Mentor: Diane Litman

TURN-TAKING CUES IN A HUMAN TUTORING CORPUS

Human conversation is a seemingly simple, everyday phenomenon that requires a complex mental

process of turn-taking, in which participants manage to yield and hold the floor with little pause in-

between speaking turns. Many linguists suggest that this is due to subtle turn-taking cues that we

subconsciously relay and process while talking with others. Identifying these cues can help improve

spoken dialogue systems, which often fail in replicating human turn-taking patterns. In this research,

turn-taking in a human tutoring corpus of spoken conversations between a teacher and a student is

examined and compared to that in other domains. Student speaking turns were divided into segments

based on pause intervals, and split into two groups: YIELD if the next segment was the tutor speaking,

and HOLD if the student continued his turn. Prosodic features were calculated for each segment, and

means were compared between the YIELD and HOLD groups. Significant differences between the two

for a feature would suggest that this feature is a turn-yielding cue; that is, it signals to the other person

that the speaker is giving up his turn to talk. Preliminary results suggest that while certain prosodic

features can be significant turn-yielding cues, this varies greatly by person.

Funding: National Science Foundation

Maxwell Garber

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Linda Jen-Jacobson

FLANKING SEQUENCE EFFECTS AND ACTIVE SITE REPULSION ARE COUPLED AT THE ECORI PROTEIN-

DNA INTERFACE

Sequence-specific protein-DNA interactions lie at the heart of most biological processes. Because of

their stringent discrimination against incorrect DNA sites, restriction endonucleases provide excellent

models for understanding the rules that govern specificity. We have previously shown that the binding

affinity of EcoRI restriction endonuclease can be dramatically affected by (1) sequences flanking the

recognition site and (2) active site repulsion (electrostatic strain) arising from the apposition of

negatively charged residues (E111, D91) to the scissile phosphate. Here we present data that reveal a

link between these two seemingly unrelated determinants of binding. Substitution of an active site

residue produces a mutant EcoRI endonuclease (E111A) which no longer shows flanking sequence

preferences; that is, the E111A protein binds cognate sites flanked by different sequences with similar

affinity. The conformational properties of flanking sequence control the precise geometry of protein-

phosphate contacts, which are structurally "coupled" to the positions of base-recognition and active-site

residues. We propose that suboptimal flanking sequence and active-site repulsion thus cause

reciprocally coupled micro-adjustments throughout the complex. Relieving strain in either the active site

(E111A) or the flanking region (best sequence) consequently relieves strain everywhere, thus optimizing

protein-DNA interactions. We do not yet know if other charge alterations (e.g. D91A) in the active site

will produce the same effects.

Funding: Samuel D. Colella Award, Department of Biological Sciences Mentoring Fellowship

Linda Glah

Neuroscience

Arts and Sciences

Faculty Mentor: Amy Wagner, MD

VOLTAMMETRIC ASSESMENT OF AMANTADINE EFFECTS ON DOPAMINE NEUROTRANSMISSION

Amantadine, typically known as a NMDA antagonist, has been used to treat Parkinson’s disease. Also,

amantadine is used as a treatment for patients with traumatic brain injury (TBI), and is often

administered in conjunction with other dopamine agents like L-dopa or methylphenidate. Although

effective in the clinical setting, amantadine’s actual mechanism of action on dopaminergic systems

remains a mystery. We use a voltammetric model to measure real time evoked dopamine overflow in

the caudate nucleus caused by stimulating neurons in the medial forebrain bundle of the rat brain. Dose

response studies have been conducted in order to evaluate characteristic changes from the baseline

level of evoked dopamine. Several doses have been studied including 10mg/kg, 20mg/kg, and 40mg/kg

(n=5) in each group. These drug effects were observed for at least 60 minutes after administration.

Results show that 10 mg/kg and 20 mg/kg have no significant impact on dopamine neurotransmission.

However, 40 mg/kg caused an averaged 70% increase in dopamine release one hour after drug

administration. Overall, amantadine can be considered a long acting dopamine enhancer with peak

changes in evoked dopamine occurring at 60 minutes post-drug administration. This dopamine

enhancement may explain clinical observations of improved cognition, mood, and motor control.

Current studies are now evaluating an animal model of TBI where a 40mg/kg dose response is compared

in injured and naïve rats.

Funding: Institute for Rehabilitation Research and NIH R21NS057348

Michael Gowen

Neuroscience

Arts and Science

Faculty Mentor: Dr. Michael Levine

ELECTROPHYSIOLOGICAL ALTERATIONS IN PREFRONTAL CORTICAL PYRAMIDAL NEURONS IN A MOUSE

WITH DEFICIENT FULL-LENGTH DISC1 PROTEIN EXPRESSION

Schizophrenia is a complex disease, the etiology of which is determined by both hereditable and

environmental factors. In a large Scottish family, a positive association between schizophrenia and a

point mutation in the DISC1 (Disrupted In Schizophrenia-1) gene was found; this mutation causes a

premature stop codon, leading to a truncated Disc1 protein. As Disc1 is a scaffolding protein involved in

neurogenesis, cell growth, and migration, its disruption may cause changes in neural circuitry leading to

the symptoms of schizophrenia. Using electrophysiological methods we recorded currents from brains

in a knock-in mouse model with a disruption in the murine homologue of DISC1 to determine the effects

of this mutation (Li et al., 2007). We recorded and compared spontaneous and evoked synaptic currents

in cortical pyramidal neurons from DISC1 mutant mice and their wild type (WT) littermates using whole-

cell voltage clamp electrophysiology. Inhibitory synaptic transmission was isolated pharmacologically

using the selective glutamate receptor antagonists NBQX and AP5, which block AMPA and NMDA

receptors, respectively, and by holding the membrane potential at +10mV. We found that the

amplitude of evoked inhibitory synaptic currents in cells from DISC1 mutant mice was larger than in WT

mice, whereas both the frequency of spontaneous synaptic currents was lower in the DISC1 mice,

relative to the control. These findings suggest that spontaneous and activity-dependent GABAergic

synaptic currents are altered; this may play a role in the expression of symptoms seen in patients with

schizophrenia.

Funding: The Amgen Foundation

Akash Goyal

Economics

Arts and Sciences

Faculty Mentor: Dr. Amy K. Wagner

MANIFESTATION AND PATHOPHYSIOLOGY OF MYELIN BASIC PROTEIN AND S100 CALCIUM BINDING

PROTEIN IN HUMAN SERUM AND CEREBROSPINAL FLUID AFTER SEVERE TRAUMATIC BRAIN INJURY

Traumatic brain injury (TBI) annually affects 2 million Americans, and 50,000 of these individuals die

each year. Such TBI causes cellular damage, which releases proteins into cerebrospinal fluid (CSF) and

serum that can be measured using Enzyme-Linked Immunosorbent Assays (ELISA). Establishing a

correlation between levels of proteins in serum and CSF can establish the brain specificity of that protein

and justify serum measurement as a part of clinical care. Previous studies have shown changes in levels

of myelin basic protein (MBP) and S100 calcium binding protein (S100B) after severe TBI thus identifying

them as potential biomarkers. With adequate transfer of the biomarkers across the blood brain barrier

after injury, serum and CSF would correlate. Using ELISA kits, the concentrations of MBP and S100B

were measured in serum and CSF of 31 and 60 TBI patients, respectively. Samples were collected from

day of injury to 5 days following injury. Concentrations were compared to protein levels from healthy

controls and were associated with various demographics to detect potential significance. Age (p<0.05),

gender (p<0.10), 6-month GOS (p<0.05) showed potential significance in certain markers. CSF and

serum levels showed preliminary correlation for S100B (r=0.185), however the population will need to

be increased in future studies to make further inferences. Using this information and respective

correlation between serum and CSF, it may be possible to better understand how serum and CSF MBP

and S100B levels correlate with one another, whether levels present in serum versus CSF are more

closely linked to outcome, and further establish the viability of these proteins as TBI biomarkers for use

in clinical care.

Funding: CDC Grant: #R49/CCR323155 and DOD Grant: #W81XWH-07-1-0701

Feng Guo

Health Promotion and Development

School of Nursing

Faculty Mentor: Denise Charron-Prochownik, PhD, RN, CPNP, FAAN

A MIXED METHOD APPROACH TO DESCRIBE THE KNOWLEDGE OF MOTHERS OF ADOLESCENT

DAUGHTERS WITH DIABETES REGARDING PRECONCEPTION COUNSELING AND PREGNANCY

Female adolescents with diabetes are at risk for reproductive-health complications. Preconception

counseling (PC) can minimize these risks. Mother’s perceptions and knowledge of PC and pregnancy

with diabetes can positively influence their daughters’ decisions/outcomes. In a previous RCT, we

examined the effects of a tailored PC program (READY-Girls) on teens with diabetes, but did not include

their mothers. The purpose of this mixed method pilot study was to describe the mothers’ qualitative

and quantitative knowledge of PC and pregnancy with diabetes. Paper-pencil questionnaires were

administered to 7 mothers of adolescent-daughters with diabetes. Qualitative data were evaluated

using content analysis with a coding scheme; 100% agreement was achieved for inter-rater reliability.

Quantitative knowledge was measured using 28-items with 2-subscales. Mothers’ mean age was 48yrs

(45-53yrs), 86% Caucasian, 86% college graduates. Top qualitative themes to knowing PC: “nothing”

(57%,n=4), general pregnancy (43%,n=3), and physician recommendations (29%,n=2). Top themes of

knowing pregnancy with diabetes: “nothing/very little” (57%,n=4), good control (43%,n=3), general

complications (43%,n=3), and infant complications (43%,n=3). Knowledge scale of PC had a mean score

of 70% (50%-86%); and pregnancy with diabetes had a mean score of 67% (50%-75%). Mothers lacked

knowledge in PC and diabetes with pregnancy. Both (mixed) methods supported these findings. Many

mothers said they knew “nothing”, or knowledge tended to be more general. Providing an educational

intervention to the mothers like READY-Girls could potentially increase their knowledge and elicit

maternal support that could help to empower their daughters to make informed decisions with

reproductive-health behaviors and prevent long-term complications.

Funding: R01 HD044097-01 from NIH-NICHD

Yeong Han

Department of Pharmaceutical Sciences

School of Pharmacy

Faculty Mentor: Yijun Huang

DATA MINING OF PDB DATABASE FOR FRAGMENT-BASED DRUG DESIGN

Data Mining of PDB Database for Fragment-based Drug Design

Fragment-based drug design is a new approach that uses X-ray crystallography or other physical

techniques to successfully screen fragment libraries for specific binding to a target protein. The

knowledge of where specific chemical fragments bind to a protein is readily available through protein

data bank (PDB) and Relibase. Querying the PDB using Relibase allow for the discovery of thousands of

small molecule fragments that bind to specific active sites of any proteins giving way to creating higher

affinity compounds. Two prominent fragments for drug-design are piperazine and para-chlorobenzyl

motifs. Chemoinformatic studies of the databases give hundreds of ligands that contain either of those

two fragments. The further investigation provides how the fragments are interacting with specific amino

acids on target proteins. This study will give the insight of new directions for drug design based on these

two fragments.

Kyle Holden

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Yong Li MD, PhD

SYNTHETIC MATRIX METALLOPROTEINASE INHIBITOR, GM6001, SLOWS MUSCLE HEALING PROCESSES

AFTER INJURY

Matrix metalloproteinases are zinc-dependent endopeptidases that aid in tissue remodeling through the

degradation of various components within the extracellular matrix. Previously, we have detected that

MMP1 is effective in overcoming the barrier of fibrosis, which can hinder successful restoration of the

damaged tissue, and ultimately lead to loss of function. To observe the mechanism behind MMP1 in the

muscle healing process, the purpose of this experiment is to investigate regenerative processes and

fibrosis formation in injured muscle through the use of the synthetic broad-spectrum MMP inhibitor,

GM6001. C2C12 myoblasts were evaluated using immunocytochemistry to assess for the presence of

stem cell markers (Sca1 and CD34) after MMP1 stimulation and after MMP1 stimulation followed by

treatment of GM6001. Also, immunocytochemistry was used to evaluate the myogenic differentiation

capabilities of C2C12 myoblasts that were either stimulated with MMP1 or treated with GM6001 after

MMP1 stimulation. Lacerations to the gastrocnemius muscle were used to represent a skeletal muscle

injury in the murine model; the areas of injury were treated with either MMP1 or GM6001 and assessed

for fibrotic scar formation. The application of the synthetic MMP inhibitor, GM6001, resulted in

increased fibrosis, decreased myogenesis, and the reduction of the percentage of cells expressing stem

cell markers within a population in vitro. These findings have implications in furthering the knowledge

of the roles of MMP1 in muscle healing and regeneration as well as providing possible insight into the

mechanisms behind these processes.

Funding: Grant of Research Advisory Committee, Children's Hospital of UPMC

Daniel Holohan

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Jeffrey Hildebrand

Designing Proteins to Spatially Regulate Actomyosin Contractility

Shroom 3 (Shrm3) is an actin-binding protein that influences epithelial morphogenesis during

development by establishing contractile actomyosin networks. Shrm3 regulates these networks by

recruiting Rho-kinase (Rock) to the tight junctions (TJs) that are located at the apical domain of the

lateral membrane, where it causes contraction by activating myosin II. To determine if Shrm3 can be

bypassed in this pathway, we sought to generate a version of Rock that is directly targeted to the TJs.

To accomplish this, we generated a chimeric protein consisting of the localization domain of Shrm3 &

the kinase domain of Rock. Individually, it was shown that Shrm3 localizes to F-actin stress fibers and

adherens junctions in epithelial cells. Furthermore, it was shown that constitutively active Rock

increases the formation of F-actin in cells, while dominant negative Rock decreases the presence of F-

actin. In future experiments, the chimeric protein construct will be transfected into epithelial cells to

see if the it directly targets the kinase domain to TJs and causes apical constriction. The morphological

changes will then be compared to those induced by the Shrm3 and Rock constructs.

Funding: Biological Sciences Department through HHMI

Christopher Horvat

Math

Arts and Sciences

Faculty Mentor: Bard Ermentrout

SPIRAL WAVES IN FLEXIBLE MEMBRANES

Rotational spiral waves are a common find in many disjoint areas of the life sciences. Especially notable

is experimental evidence for their existence in both heart and brain tissue. For modeling these organs, it

is often convenient to use the mathematical study of flexible membranes. Here, we present that spiral

wave formation is evident for even small-scale specific flexible membrane problems, using a number of

different models.

Funding: NSF RTG

Jennifer Huling

Bioengineering

Swanson School of Engineering

Faculty Mentor: Dr. Frank Witte

THE DEVELOPMENT OF A HISTOCHEMICAL STAINING METHOD FOR DETECTING

ACETYLCHOLINESTERASE IN SOFT TISSUE, BONE AND CULTURED CELLS

Magnesium metal offers a viable option for a biocompatible and biodegradable material. In bone, the

corrosion of magnesium causes temporarily increased bone growth by increasing osteoblast activity. The

enzyme acetylcholinesterase (AChE), typically thought to be involved with cholinergic synapse function,

has been found to play a role in osteoblast function. The goal is to be able to monitor AChE as an

indicator of osteoblast behaviour and a measure of the change induced in bone because of magnesium

corrosion. To develop a staining method for the detection of AChE, soft tissue and bone samples, as well

as cultured cells, were stained using the partially established Karnovsky and Roots method and

immunohistochemical methods. Karnovsky and Roots staining was found to only work on frozen

sections of tissue. The mouse-anti-AChE clone ZR3 antibody was found to be ineffective as a primary

antibody, but the clone HR2 was found to work with both enzyme and fluorescent staining methods in

frozen sections and those embedded in paraffin and plastic. Further studies can apply the developed

method of positively identifying AChE in bone to help determine AChE’s role in osetoblast behaviour and

how magnesium corrosion stimulates increased bone growth.

Funding: Hannover Medical School

Alex Ireland

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Jeff Brodsky

The Identification of Small Molecule Polyomavirus Inhibitors Based on a Novel Scaffold

Polyomaviruses are common viral agents in the human population, and can become pathogenic in

immunosuppressed populations. Because most treatments non-specifically target DNA replication, no

treatments for polyomavirus infection without undesirable side-effects are currently available.

However, all polyomaviruses express a Large Tumor Antigen (T Ag), which is a multi-domain, 708 amino

acid protein. The replication of SV40 (Simian Virus 40), a member of the Polyomaviridae family, requires

two domains within the viral-encoded T Ag: The ATPase domain, which serves as a helicase for viral

DNA, and the N-terminal J domain, which stimulates the activity of host-encoded Hsp70 molecular

chaperones. Previous screening of pyrimidinone-peptoid hybrid compounds in our laboratories

identified MAL2-11B as an inhibitor of viral replication, Hsp70 stimulation, and T Ag ATP hydrolysis (IC50

&#8776; 50 &#956;M). Therefore, we proposed that the MAL2-11B scaffold might be co-opted to

identify more effective inhibitors of T Ag ATPase activity and viral replication. We now report that a

MAL2-11B tetrazol derivative inhibits T Ag ATP hydrolysis (IC50 &#8776; 20 &#956;M). The same

derivative also displays significantly more potent inhibition of T Ag-mediated Hsp70 ATP hydrolysis than

MAL2-11B. Moreover, the tetrazol is more effective at inhibiting the replication of viral DNA in SV40-

infected cells, possibly because the derivative is anticipated to possess a greater ability to permeate the

membrane than MAL2-11B. We are currently screening additional tetrazol-based MAL2-11B derivatives

to elucidate a structure-activity relationship. We suggest that these may provide a specific, novel

therapeutic treatment for polyomavirus infection and related disease.

Funding: Beckman Scholars Program

Helmet Karim

Health Information Management (HIM)

School of Health and Rehabilitation Services (SHRS)

Faculty Mentor: Leming Zhou

MANUALLY ANNOTATING GENES IN MULTIPLE DROSOPHILA GENOMES USING EVIDENCE-BASED

APPROACH

As sequencing technology improves, we have access to an increased amount of fully sequenced

genomes. The first and most important step in analyzing these genomes is gene annotation, that is,

accurately identifying the locations of genes along the genome and determining the function of these

genes. After annotations of genes within a species genome, many further analyses can be performed to

obtain more meaningful understanding of the genome. In this work, we have manually annotated

sequences from a few Drosophila species (D. erecta, D. majovensis, D. grimshawi) using strictly

computational methods and an evidence-based approach. This particular organism is selected because

Drosophila species are extensively used in basic science and biomedical research, twelve genomes of

Drosophila species have been fully sequenced, and the genome of Drosophila melanogaster is well

annotated. This manual gene annotation project utilizes ab initio gene finding programs, the accurate D.

melanogaster gene models, the BLAST search programs, and the UCSC genome browser to determine

the precise locations of each coding exon in genomic sequences of multiple Drosophila species. The

obtained gene models for these species are then compared to their ortholog genes in D. melanogaster

to determine the function of these genes and gene exon-intron structure changes. This comparison can

also show sequence divergence of multiple Drosophila species at different evolutionary distances in

both coding and non-coding regions. The obtained results in this project have been submitted to a

central database and more data will be accumulated to perform further analysis.

Fudning: NSF IIS-0938393 and a funding from HHMI.

Joseph Kaus

Chemistry

Arts and Sciences

Faculty Mentor: Dr. Lillian Chong

EFFICIENT EXPLICIT-SOLVENT MOLECULAR DYNAMICS SIMULATIONS OF MODEL MOLECULAR

RECOGNITION SYSTEMS

Protein binding is a rare event because a long time is spent waiting for the event to occur, rather than

the duration of the event itself. This makes it difficult to study using standard molecular dynamics

simulations. The goal of this research is to develop a computational approach for simulating these rare

events. This approach is known as the weighted ensemble (WE) approach. This technique enhances the

efficiency of computer simulations of rare events. It uses computer time more efficiently by spending

less time simulating molecules that are not likely to bind, and more time simulating molecules that are

likely to bind. It does this by tracking how close the molecules are to binding, making copies of the

simulations that are closer to binding, and terminating simulations that are farther from binding. Each

simulation is assigned a statistical weight, so making copies of a simulation or terminating simulations

does not bias the results. This technique gives the pathways of binding (which is a movie of all of the

different ways the molecules bind), as well as the relative rate at which the molecules bind. Here we

compare the results and efficiency of WE approach to that of standard molecular dynamics simulations

on four model systems. Our results reveal that the WE approach is at least four times more efficient at

sampling these molecular association events.

Funding: NSF CAREER; Brackenridge Fellowship; Office of Experiential Learning

Matthew Keddie

Chemistry

Arts and Sciences

Faculty Mentor: Alexander Star

CARBON NANOCAPSULES: TAILORED LOADING THROUGH SYNTHETIC CONTROL

The synthesis of nitrogen-doped carbon nanotube cups through chemical vapor deposition (CVD) has

created an intriguing form of multi-walled carbon nanostructure. By performing CVD with metal catalyst,

ethanol, and a nitrogen source of ammonia, individual segments are synthesized in a “stacked cup”

conformation. We have observed using scanning electron microscopy (SEM), transmission electron

microscopy (TEM), and atomic force microscopy (AFM) the diameters of such architectures are

contingent upon metal catalyst diameter. In addition, we have discovered individual segment length is

highly dependent upon nitrogen concentration. Based on this work, we have tailored the volumetric

parameters of these nanostructures. Furthermore, separation of size-controlled segments is promoted

via mechanical grinding, where individual cups are isolated from their fibrous shell. Two individual cups

can be cross-linked using glutaraldehyde to form a nanocapsule, upon which particles can be enclosed.

The capsule lengths are highly dependent on the quantity of particles that can be contained. Based on

their size, shape, and inherent hollow interior cavity, these materials possess limitless potential to be

utilized as media for energy storage and biomedicine. This methodology serves as a foundational

understanding of employing an engineered system as an eventual segue into the realm of drug delivery.

Funding: The National Energy Technology Laboratory

Devin Knisely

Acute Tertiary Care

School of Nursing

Faculty Mentor: Dr. Paula Sherwood

LONGITUDINAL PATTERNS OF DEPRESSION IN PATIENTS FOLLOWING ANEURYSMAL SUBARACHNOID

HEMORRHAGE

Depression is common following aneurysmal subarachnoid hemorrhage (aSAH). Most studies use

population-average analyses masking identification of distinct groups/patterns of change over time;

data vital to designing and implementing interventions. This analysis used group-based trajectory

analysis with data from persons with aSAH to determine patterns of depressive symptoms over the year

following hemorrhage. Data from adult patients (n=193) with a moderate to large aSAH and no pre-

existing neurologic disease were recruited at insult (R01–NR001339). Depressive symptoms (Beck

Depression Inventory II) were assessed during face-to-face interviews at 3 and 12-months following

hemorrhage. Group-based trajectory analysis (SAS) was conducted using clinical/statistical criteria to

determine number and shape of trajectory groups. Analysis identified two distinct linear trajectories.

Most participants (75%, n=145) had low levels of depressive symptoms at 3-months (mean=6.46,

SE=1.1), scoring below the established cutoff for being at high risk for clinical depression. One-quarter

(25%, n=48) were at moderate risk for depression at 3-months following hemorrhage (mean=20.56,

SE=1.95). Neither mean significantly changed from 3 to 12-months following hemorrhage (p=0.86, 0.70

respectively). Data suggest two distinct patterns of depressive symptoms exist. One-quarter of the aSAH

population may be at moderate risk for depression, remaining unchanged at 12-months. Future

analyses should identify characteristics of each group to allow targeted interventions.

Funding: NIH R01 NR004339

Melissa Knox

Psychology

Arts and Sciences

Faculty Mentor: Judith A. Erlen

DIARY USE AND MEDICATION ADHERENCE IN CONTROL SUBJECTS

Diary keeping of medication taking can assist self-monitoring, often a component of adherence

interventions. The purpose of this study is to examine diary use and medication adherence in a control

arm of a randomized study. This sub-study of 97 subjects from a control condition of a randomized

clinical trial on behavioral interventions to improve HIV medication adherence examines the use of

diaries and their relationship to changes in medication adherence. Seventy-eight subjects recorded their

medication taking using paper diaries over 18 months. Adherence was measured with a self-report

medication adherence scale (SMAS) and electronic event monitoring (EEM) recorded at baseline and 18

months. Open ended exit interviews assessing self-perception of improved adherence were completed

after final follow-up at 18 months (n=74). Change in adherence for each measure was categorized as

either 1) improved or 2) maintained or diminished. Findings demonstrate no relationship between diary

keeping and adherence. While participants reported improved adherence during the study with the

SMAS (t=1.758, p=.083), EEM indicated a decrease in adherence (t=-2.404, p=.018). Neither measure

related to diary use (SMAS: &#935;²=1.557, p=.212; EEM: &#935;²=.084, p=.772). On interview, 39 of the

control subjects reported improved perceived adherence from study participation; this also was not

related to diary use (&#935;²=.056, p=.813). Diary keeping was unrelated to medication adherence in

the control group in this clinical trial suggesting that diary use was not an intervention. Because of

subject perception, future medication adherence studies should examine the role of diary keeping on

control subjects.

Funding: NIH, NINR 1R01 NR04749

Belinda Lao

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Lydia Daniels

LEPTIN IN ASSOCIATION WITH NEUROINFLAMMATION AND OUTCOME AFTER SEVERE TRAUMATIC

BRAIN INJURY (TBI)

The hormone leptin is usually recognized for its role in metabolic regulation and energy balance.

However, leptin has been shown to exhibit neuroprotective and anti-apoptotic functions in the human

brain, as well as a potential role in the formation and modulation of the immune response. This study

investigates the cerebrospinal fluid (CSF) levels of leptin in association with pro- and anti-inflammatory

markers and markers of cell adhesion (sICAM-1 and sVCAM1) in severe adult traumatic brain injury (TBI)

patients to determine trends in relative marker levels for informing prognosis and effective

management of the inflammatory response following TBI. CSF was collected from the 14 patients within

an acute period of 5 days (144 hours) after the TBI, through an EVD (extraventricular drain) every 12

hours. Eleven healthy control samples were obtained from volunteers who met the established criteria

for health and demographic. The leptin concentration was tested using ELISA (Enzyme-linked

immunosorbent assay), while cytokines (IL-1, IL-6, IL-8, IL-10, TNFa), sVCAM-1 and sICAM-1 were

measured by xMap Luminex technology. Patient medical records were utilized to document incidence

of sepsis and other complications in TBI patients. The aim of the study is to describe the relation

between leptin and the formation of immune response in TBI patients while controlling for factors like

septic complications and demographic background.

Funding: DODW81XWH-071-0701

Naomi Latorraca

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Michael Grabe

MEMBRANE BENDING STABILIZES TRANSMEMBRANE PROTEINS CONTAINING CHARGED RESIDUES

The lipid bilayer of cellular and organellar membranes deforms to stabilize the inclusion of

transmembrane (TM) proteins. Previous studies have demonstrated that membrane bending helps to

expose proteins’ charged residues to the polar extracellular environment and phospholipid head

groups. The present study uses a computer search algorithm to generate a membrane boundary curve

that most stably accommodates the inclusion of a simple TM &#61537;-helix. The algorithm then uses

continuum electrostatics, which assigns constant dielectric values to the extracellular and bilayer

environments, to calculate the peptide’s minimum free energy of insertion into the bilayer based on the

membrane’s configuration. Peptides used in these simulations consisted primarily of hydrophobic

residues, with a central position containing one of each of the twenty amino acids. The individual

insertion energies obtained for these peptides permitted the construction of a biological hydrophobicity

scale, which ranks the set of amino acids in terms of their individual apparent free energies of insertion.

The data compare favorably with existing hydrophobicity scales, falling within range of both

experimental and computational values. Additionally, the algorithm was tested on other simple

peptides, including those with multiple charged residues. This novel method demonstrates the

sensitivity of the membrane's ability to find energetically favorable accommodations for proteins

positioned at the water-lipid interface, providing a computationally efficient way to model essential

aspects of protein-membrane interactions.

Funding: Howard Hughes Medical Institute 2010 Summer Undergraduate Fellowship to N.L. and NSF

CAREER Award to M.G.

Cory Leeson

Bioengineering

Swansons School of Engineering

Faculty Mentor: Dr. William Wagner

SPATIAL CONTROL OF GENE EXPRESSION IN A POLYMERIC SCAFFOLD

Tissue engineering and regenerative medicine are promising approaches to restore healthy tissue after

injury – such as myocardial infarction. One important approach to encourage regeneration is to create

healthy heart muscle in vitro for implantation in the place of damaged myocardium. Although

controlling cell behavior within a scaffold is a challenge, such control could allow the generation of

complex tissue constructs. Control can be achieved by inducing expression of desired genes. Inducer

molecules, such as RheoSwitch Ligand 1 (RSL1), can provide control of target genes in cells transfected

with the corresponding gene expression system. In this project, a biodegradable elastomer important

for tissue engineering was utilized to construct a highly porous scaffold with defined regions where RSL1

was either present or absent. Cells grown on this scaffold expressed the reporter green fluorescent

protein (GFP) predominately in regions of the scaffold that contained RSL1.The normalized green/blue

ratio (GFP to blue nuclear stain) of image intensity data across the center of the scaffold confirmed a

localized control of GFP expression. The ability to spatially control gene expression within a cell seeded

scaffold may open opportunities to drive cell behavior toward the generation of complex tissue

constructs.

Funding: National Institute of Health

Christopher Lippert

Mechanical Engineering

Swanson School of Engineering

Faculty Mentor: Dr. Gregory Reed

Sample Survey of Smart Grid Approaches and Technology Gap Analysis

Research and development in the field of “smart grids” is advancing at an ever expanding rate, with an

increasing number of industry participants and other key constituents internationally, including

government entities and educational institutions. This survey was undertaken with the intent of

determining representative approaches from various participants, and combining them into an

overarching view of the industry as a whole. As a result, the more practical and efficient methods of

improving the electrical grid were revealed, as well as gaps within the existing technology and

standards. The most apparent gaps were in the following main areas: common communications;

improved transmission and distribution controls; real-time information and incentives for both the end-

user and the utility; self-healing grids; energy storage and renewable integration; and improved

standards for the industry. In particular, our research group has begun development of a computer

model to simulate the performance of energy storage for load management on a distribution substation.

Using the loading data from a substation, it will model the effects of installing various levels of energy

storage on that substation. This will allow for the determination of ideal energy storage sizes for a

particular substation, which will be essential to the economic implementation of the smart grid.

Funding: Mascaro Center for Sustainable Innovation (MCSI)

Hei Ma

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Zhou Wang

GENETIC ANALYSIS OF FACTORS INFLUENCING ANDROGEN RECEPTOR LOCALIZATION USING YEAST AS

A MODEL SYSTEM

Androgen receptor (AR) is a key molecule in the development and progression of prostate cancer, which

is a major disease among men in the United States. In normal prostate, AR regulates androgen response

genes expression in a ligand-dependent manner. In the absence of ligand, AR is localized in the

cytoplasm and is transcriptionally inactive; however, it is translocated into the nucleus and becomes

active after ligand binding. It has been shown that in castration resistant prostate cancer, AR remains in

the nucleus regardless of ligand availability. The mechanism of how this occurs remains unknown. In

order to gain more insight into the pathway mediating AR intracellular trafficking, we seek to identify

molecular factors that influence this process using S. cerevisiae (budding yeast) as a model system due

to its genetic tractability. Previous work in our lab has identified a novel nuclear export signal (NES-AR)

in AR that is necessary and sufficient for AR cytoplasmic localization. Yeast expressing a fusion protein

containing GFP, NLS-SV40, and NES-AR was subjected to transposon mutagenesis and the localization of

such protein was examined under fluorescence microscopy. Mutants defective in mediating cytoplasmic

localization of the fusion protein will be identified. The results from this study will give us a better

understanding of AR localization pathway in hopes of developing treatments that modulate AR activity

in humans to prevent or delay prostate cancer progression.

Funding: NIH R37 DK51193 and R01 CA 108675

Catherine Madden

Chemistry

Arts and Sciences

Faculty Mentor: Christopher Matranga, National Energy Technology Laboratory

SYNTHESIS AND CHARACTERIZATION OF A SERIES OF PORE FUNCTIONALIZED PILLARED-LAYERED

METAL ORGANIC FRAMEWORK TYPE MATERIALS FOR CO2 CAPTURE

Pillared-layered compounds are an exciting material which show promise as CO2 capture agents. They

belong to a general class of compounds called Porous Coordination Polymers (PCPs) which are formed

by reacting transition metal ions with various organic linker molecules. The chemistry used to synthesis

these compounds creates a material whose pore size, network type, and chemical makeup is controlled

in a directed and rational manner by the choice of metal ions and organic linkers. In this regard,

pillared-layered compounds are closely related to metal organic frameworks (MOFs), but typically have

distinct performance and stability advantages over traditional MOFs. One of the largest obstacles in

making new PCPs for CO2 applications is creating the organic linkers needed for the pore walls of the

structure. The addition of different functional side groups on these linkers allows one to modulate the

chemical affinity of the PCP pore wall towards CO2 or other impurity gases in the flue of a coal fired

power plant. To this effort, several novel functionalized bi-pyridyl derivatives containing -NH2, -NO2, -

CH3, -F, -C(O)H, -C(O)OH, and -OCH3 groups were synthesized by either an amidification reaction or the

Suzuki coupling reaction. These linkers were then incorporated into a pillared-layered material to create

new pore- functionalized PCPs. Each of these structural analogs was evaluated for CO2 capture and

separation applications using isotherm and porosimetry techniques. From this work, the systematic

effect of adding these functional groups to the bi-pyridyl linkers was assessed and will be used to

optimize these CO2 capture materials.

Funding: Mickey Leland Energy Fellowship (Department of Energy)

Michael Mihalevic

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Jeffrey Brodsky

CHARACTERIZATION OF THE ROLE OF SMALL UBIQUITIN-LIKE MODIFIER (SUMO) IN CELLULAR QUALITY

CONTROL

Molecular modifiers function to confer new properties upon their target proteins. For example, binding

of a particular modifier can either activate or deactivate a protein, or even change its distribution in the

cell. An interesting attribute of these molecules is the impact that they may have on protein stability and

cellular quality control in response to cell stress. Upon binding of molecular modifiers a protein could be

protected from, or targeted for, degradation. One prominent molecular modifier is ubiquitin; its binding

to a protein can act as a signal for proteasome-mediated degradation. We propose that Small Ubiquitin-

like Modifier (SUMO) is involved in the cellular stress response. In our experiments we subjected cells

with mutations in the SUMOylation pathway to assays that measure various types of cellular stress. We

observed that the strains were vulnerable to exposure to the reducing agent dithiothreitol (DTT), which

unfolds proteins in the Endoplasmic Reticulum, as well as chemicals that induce general protein

misfolding and osmotic stress. Additionally, we performed cycloheximide chase assays to evaluate if the

degradation of specific substrates in these mutant backgrounds was altered. Results of the experiments

revealed that deSUMOylation plays an important role in the degradation of a model misfolded substrate

CPY*. Future studies aim to elucidate the function of SUMO in protein degradation. For example,

cycloheximide chases and immunoprecipitation assays will reveal if CPY* is SUMOylated or if the defect

degradation occurs indirectly.

Funding: Chancellor's Undergraduate Research Fellowship

Kaitlin Mitchell

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Graham Hatfull

IDENTIFYING MYCOBACTERIOPHAGE GENES CYTOTOXIC TO MYCOBACTERIA

Bacteriophages, viruses of bacteria, have evolved to effectively take over and utilize the cellular

machinery of their host. While the major processes of phage infection are well known, we sought to

identify single phage gene products that interact with host proteins to inhibit cell growth. Genes of

unknown function from mycobacteriophage Giles were chosen for screening with a cytotoxicity assay.

Giles was chosen for this experiment because there is a large amount of information known about its

gene requirements and expression acquired through work in our lab. Individual genes were cloned into a

nitrile-inducible vector system, allowing for controlled gene expression. Twenty-four constructs were

made and transformed into Mycobacterium smegmatis mc2155, a non-pathogenic relative of M.

tuberculosis, and are being tested for their effect on cell survival. The LysA gene from phage L5 was used

as a positive control, as it encodes a protein shown to lyse cells. Genes identified as cytotoxic will then

be tested on M. tuberculosis. The study of such genes and the interactions of their gene products with

host proteins will allow us to discover potential drug targets and aid in future development of anti-

tuberculosis drugs.

Funding: HHMI

Leah Nissle

Biological Science

Arts and Sciences

Faculty Mentor: Arundhati Ghosh, Ph.D

TEACHING TECHNIQUES IN UPPER-LEVEL UNDERGRADUATE ANIMAL PHYSIOLOGY COURSE

A pilot study was done in an upper-level undergraduate Animal Physiology course in order to determine

the effectiveness of various teaching techniques. Students were mostly juniors and seniors, preparing

for Medical, Dental, Veterinary Schools or Graduate schools. Students were given two types of

worksheets: one with direct questions purely from the textbook and lectures, the other with a case-

based questions (having to do with a real life situation or in association with the organ system studied

that week in the lecture). After completing each worksheet, students were surveyed about their

thoughts on if the assignment helped them retain and understand the information for the exam, and if

the assignments prepared them for a real life situation or a patient, among other things. Many students

felt that the text-based questions were effective in helping them prepare for an exam, but that the

questions did not prepare them for a real life situation. More students felt that the case-based lessons

were effective or very effective than were the text-based questions. Based on the students' opinions,

the case-based lessons were more successful in teaching the topics on the exam. Case-based studies

have prepared them better for the professional schools.

Alexis Nolfi

Bioengineering, Psychology

Swanson School of Engineering, Arts and Sciences

Faculty Mentor: Steven Abramowitch, PhD

BONE TISSUE ENGINEERING AS A SCIENCE AND MATH LEARNING MODEL FOR YOUTH OUTREACH

EDUCATION

The Pittsburgh Tissue Engineering Initiative (PTEI) has created an outreach program to encourage middle

and high school students to pursue continued education in the science and math fields. PTEI’s program

in Pittsburgh, “A Starfish Can Grow A New Arm, Why Can’t I?”, and North Carolina A&T State University’s

program “Nano-to-Bio Summer Camp” have allowed approximately seventy students to develop

quantitative, analytical and technical skills in a simulated laboratory environment. Based on current

research, modules were created to engage and involve students in tissue engineering (TE) activities.

Original bone tissue engineering modules were established to teach the major components of the TE

triangle including signals, scaffolds, cells and blood. One of the main objectives was to teach the

students the properties and materials of a tissue engineered scaffold. Students created a scaffold using

poly(lactic acid) and performed an assay exploring the toxicity of various scaffold degradation products.

Campers were taught the importance of adding signals such as growth factors to scaffolds in order to

promote bone formation. Another main objective was to explore the mechanical properties of artificial

materials such as joints and blood. Lastly, students explored the structural properties of bones and how

these properties are used to design bone grafts in regenerative medicine. Students were required to

take pre and post tests evaluating their knowledge of tissue engineering. These tests indicated a

mastery of basic tissue engineering principles. This confirmed that the new and previously used

modules were relevant and exciting to students. Future work will include improvements to the activities

and development of new activities based on student feedback and integrating the modules in middle

and high school classes. New models may also be developed to incorporate additional subject matter.

Funding: University of Pittsburgh BioE REU Summer Program

Amenawon Ogiefo

Biological Sciences

Arts and Sciences

Faculty Mentor: Graham Hatfull and Deborah Jacobs-Sera

The Microbiological Chacterization of Mycobacteriophage DeadP

Bacteriophages are viruses that infect bacterial hosts. They are the most common and diverse organisms

on Earth, and collectively comprise the single largest pool of genetic information. Mycobacteriophages

are important because of their ability to infect the mycobacteria, the most notable of the genus being

Mycobacterium tuberculosis, the causative agent of human tuberculosis. Mycobacteriophage DeadP

was isolated from a soil sample in the South Oakland region of Pittsburgh, PA. Mycobacteriophage

DeadP forms small, turbid plaques with a medium-sized ‘halo’ on a lawn of M. smegmatis, a fast-

growing, non-pathogenic relative of M. tuberculosis. This study describes the isolation, purification and

microbiological characterization of Mycobacteriophage DeadP through electron microscopy and

restriction digest analysis, as we prepare to send DeadP for 454 pyrosequencing.

Funding: Howard Hughes Medical Institute

Yewande Olugbade

Biological Sciences

Arts and Science

Faculty Mentor: Dr. Anthony Kline

PERSISTENT DELETERIOUS EFFECTS ON SPATIAL LEARNING FOLLOWING CHRONIC ADMINISTRATION

OF HALOPERIDOL OR RISPERIDONE AFTER EXPERIMENTAL TRAUMATIC BRAIN INJURY

Antipsychotic drugs (APDs) are provided to alleviate clinical traumatic brain injury (TBI)-induced

agitation, albeit the effects on functional recovery remain unclear. Previous studies have shown that

haloperidol (HAL) and risperidone (RIS) impair motor/cognitive function. Still unknown is how long the

adverse effects induced by these APDs remain after their cessation. To address this issue, fifty-nine

anesthetized male rats received a cortical impact or sham injury and then were randomly assigned to

four treatments (HAL 0.5 mg/kg, RIS 0.45 mg/kg, vehicle; VEH 1 mL/kg, or bromocriptine; BRO 5.0

mg/kg; a positive control for D2-receptor action) that began 24-hours after surgery and were

administered i.p. every day for 19-days. Motor/cognitive tests were conducted on days 1-5/14-19.

Treatments were discontinued on day-20 and the rats were re-evaluated at 1 and 3 months. During the

treatment phase, both APDs significantly impaired cognition vs. VEH and BRO. Furthermore, BRO was

better than VEH (p=0.0042). At 1-month, both APDs still exhibited significant cognitive impairment vs.

VEH and BRO. At 3-months, only the HAL group was still impaired vs. VEH, but both APDs continued to

be deficient vs. BRO. No differences were revealed between the APD groups. Motor deficits/recovery

did not differ among groups. These data replicate previous reports of HAL and RIS impeding cognition

after TBI and extend those results by showing that the deleterious effects persist 3 months after

discontinuation. The benefits observed with BRO suggest that the adverse effect of the APDs is

mediated by D2-receptor antagonism. Evaluation of non-D2-antagonist APDs is needed.

Daniel Perchy

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Huinan Liu

MAGNESIUM ALLOYS AS NEXT-GENERATION BIODEGRADABLE MEDICAL IMPLANTS

Biodegradable magnesium implants may offer a viable alternative to traditional surgical treatments for

torn tendons and ligaments. Magnesium can be implanted at the site of injury and then degrade

benignly over time. As the implant degrades, the injured tendon or ligament can re-grow by cell

proliferation and extracellular matrix production, eventually restoring tissue function. In order to

optimize the growth of connective tissue cells on magnesium, the effects of surface microstructure and

chemical environment on implant degradation and cell function must be studied. It is important to

understand how cells would grow and differentiate on magnesium with various properties. My

experiments investigated the degradation of pure and yttrium-alloyed magnesium samples over time.

Samples were immersed in either water, phosphate buffered saline solution, cell medium, or cell

medium supplemented with fetal bovine serum to study their degradation in different environments.

The change of weight and surface appearance of magnesium and magnesium-yttrium alloy samples was

measured and monitored until the samples were fully degraded. The pH variation of immersion

solutions was tracked to understand how the degradation of magnesium affects the local physiological

environment. This study provides a basis for the design of magnesium alloys as next generation

biodegradable medical implants .

Funding: NIH, University of Pittsburgh

Nicholas Perri

Chemistry

Arts and Sciences

Faculty Mentor: Tara Meyer

ELECTROPLASTIC ELASTOMER DEVELOPMENT: QUANTIFICATION OF IRON ION CROSSLINKS

Electroplastic elastomers, materials whose stiffness can be altered by electrochemical stimulation,

should prove valuable for a wide range of applications. Using hydrogel copolymers of 4-

polystyrenesulfonic acid and sodium acrylate we have developed a composite material that can be

reversibly converted between a soft and a hard state by the reduction or oxidation of metal ion

(Fe2+/Fe3+) crosslinks. In order to optimize control and to verify reproducibility, it is important to

quantify the number of crosslinking metal ions. We have therefore developed an analysis method to

determine the total iron content of our electroplastic elastomers. Cross-linked hydrogels were treated

with concentrated hydrochloric acid in order to break down the polymer network and release the Fe3+

bound to the monomers. By exploiting the characteristic peak for Fe3+ at 362 nm, the quantity of iron

in the sample was determined by UV-Vis spectroscopy. Control studies were conducted to verify the

accuracy of the method.

Funding: National Science Foundation

Lindsay Plavchak

Biological Sciences

School of Arts and Sciences

Faculty Mentor: Dr. Jeffrey Brodsky

CHARACTERIZING THE ROLE OF MOLECULAR CHAPERONES IN THE DEGRADATION OF THE EPITHELIAL

SODIUM CHANNEL

The Epithelial Sodium Channel (ENaC) is an ion channel found in the epithelial membrane of both the

kidney and the lungs, where it tightly regulates sodium reabsorption within the body. Because salt and

water homeostasis is of such critical importance, even slight errors in ENaC function can lead to major

health concerns such as hypertension, hypotension, pulmonary edema, and Cystic Fibrosis-like

symptoms. ENaC is composed of three homologous subunits – &#945;, &#946;, and &#947; – that

assemble in the ER to form the mature ENaC protein. However, formation is highly inefficient,

oftentimes leading otherwise functional subunits to be degraded through a process known as

Endoplasmic Reticulum Associated Degradation (ERAD). To determine how ENaC is degraded, we have

made a yeast expression system. Previous results established that Jem1 and Scj1, two ER lumenal Hsp40

chaperones, play a role in ENaC degradation. Jem1 and Scj1 facilitate ATP hydrolysis by the ER lumenal

Hsp70 chaperone, BiP. Furthermore, we found that Lhs1 and Sil1, two nucleotide exchange factors

(NEFs) for BiP, may play a role in ENaC subunit degradation. These results led to the hypothesis that the

Hsp40s and NEFs work in combination to facilitate ENaC degradation. To test this hypothesis, the extent

of &#945;ENaC degradation was assayed in a newly created lhs1&#8710;jem1&#8710;scj1&#8710;

strain. Results reveal that the combined effect of deleting Lhs1, Jem1, and Scj1 further stabilizes ENaC in

comparison to the single deletions, indicating a synthetic effect. Consistent with these data, the

lhs1&#8710;jem1&#8710;scj1&#8710; deletion is synergistically sensitive to agents that induce ER

stress.

Funding: Howard Hughes Medical Institute Academic Year Undergraduate Research Fellowship;

American Heart Association Summer Undergraduate Research Fellowship

Christopher Rovensky

Chemical and Petroleum Engineering

Swanson School of Engineering

Faculty Mentor: Dr. Amy E Landis

BIOFUELS ON MARGINAL LANDS

Biofuel-based phytoremediation is being evaluated as a beneficial means to repurpose potentially

contaminated underutilized urban marginal lands. Biofuel crops, such as the canola, corn, soybean, and

sunflowers studied, are able to accumulate high concentrations of heavy metals that typically

contaminate soil in brown- and greyfields. Students have been traveling to sites shared through the

generosity of a local Pittsburgh nonprofit community group for the past three years to collect soil

samples before planting and shortly after harvesting occurred. Soil samples collected from marginal

lands throughout Pittsburgh were dried, ground, and digested using acid digestion. Soluble digested

samples were analyzed with atomic absorption spectroscopy (AAS) to determine the heavy metal

concentrations for 9 metals. Heavy metal concentration data has been collected for several sites since

2008, and new sites have been identified for the 2009 and 2010 growing seasons. Furthermore, the

team collaborated with ALCOA and Pennsylvania Department of Environmental Protection researchers

to determine the ability of plants to remediate contaminated and highly acidic mine refuse. A

greenhouse was constructed at the University of Pittsburgh, and preliminary tests are under way using

mine refuse provided by ALCOA and crops grown by the University of Pittsburgh team of researchers.

The biofuel crops could improve soil conditions at the sites, as well as alleviate the economic and social

blight associated with vacant urban lands.

Funding: NSF CBET Award #0933249 and NSF MCSI REU Program

Andrew Savinov

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Graham F. Hatfull

INVESTIGATION OF THE PUTATIVE PHOSPHOESTERASE ACTIVITY OF BXB1 GP47

Bxb1 is a temperate bacteriophage that infects and forms lysogens of Mycobacterium smegmatis. The

Bxb1 lysogeny system depends on site-specific DNA recombination catalyzed by Bxb1 serine-integrase.

Bxb1 gp47 protein, the Recombination Directionality Factor (RDF), alters integrase site-specificity to

promote excision and inhibit integration. Bxb1 gp47 is an unusual RDF with no functionally

characterized homologs, although 15 other mycobacteriophages encode closely related proteins. The

presence of a homolog in phage L5 in particular suggests a secondary role for this protein, because L5

encodes a tyrosine-integrase and the known requirements for recombination do not include the Bxb1

gp47 homolog L5 gp54. Our prior BRED phage mutagenesis work showed that gp47 indeed has a

secondary role in vivo (in lytic propagation of Bxb1) and that this secondary role is separable from the

excision-promotion activity. L5 gp54 and Bxb1 gp47 align well with the consensus sequence for the

calcineurin-like phosphoesterase domain, which consists of four main motifs. We are investigating the

hypothesis that Bxb1 gp47 has a phosphoesterase activity. We have purified His-tagged variants of wild-

type gp47 and two phosphoesterase motif mutants; purification of functional proteins was tested by

RDF activity assays. Preliminary phosphatase assays showed no activity from wild-type gp47 in the

standard reaction buffer. Further assays are now underway with various divalent metal ions added, as

calcineurin-like phosphoesterases are predicted to be metal-dependent. If the putative phosphoesterase

activity and dependence on intact motifs is confirmed, further work will focus on correlating these

results with the lytic-cycle viability of motif mutant Bxb1 phages.

Funding: US Steel Foundation Undergraduate Research Award (Arts and Sciences Research Award, to AS)

Amy Scarbrough

Biological Sciences

Computer Science

Arts and Sciences

Faculty Mentor: Dr. Michael Grabe

WEIGHTED ENSEMBLE SIMULATION OF ALTERNATING ACCESS IN SODIUM-HYDANTOIN TRANSPORTER

MHP1

Many transport proteins use the energy in sodium gradients to drive the uptake of small molecules.

These proteins perform functions as diverse as the withdrawal of neurotransmitters from the synaptic

cleft, the removal of sugar from the gut, and the salvage of nucleobases and related metabolites. It is

thought they operate via an alternating access mechanism in which substrate is bound in an outward-

facing conformation followed by a transition to an inward state that delivers the cargo to the cell. X-ray

structures of both states exist, but the long time required to see such slow events makes studying this

transition using traditional molecular simulations prohibitive. We employed the weighted ensemble

method, which focuses computational efforts on rare events, to study transitions in the benzyl-

hydantoin transporter Mhp1. With this method, which we accelerated using the computational

efficiency of graphics processing units, we have observed thousands of transitions. Our analysis of these

trajectories using root mean squared deviation calculations and techniques such as protein

fingerprinting reveals insights into the mechanism of alternating access, which we believe will impact

our understanding of the operation of these proteins and their roles in human diseases.

Funding: Department of Biological Sciences, through HHMI

Lauren Schmidt

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Graham Hatfull

CHARACTERIZING THE MYCOBACTERIOPHAGE DAVINCI PARTITIONING GENES: SIMILARITIES TO

MYCOBACTERIOPHAGE REDROCK AND BACTERIAL HOMOLOGUES

Many bacteria and plasmids encode partitioning proteins that are essential for normal chromosomal

segregation during replication. There are three essential components of these partitioning systems:

parS, a cis-acting regulatory site, and parA, and parB, both protein-encoding genes. Some

bacteriophages also encode these proteins to ensure stability of the extrachromosomal lysogenic state.

The E. coli phage P1 has long been known to have a partitioning system, but only recently has this

system been discovered in mycobacteriophages. Only five sequenced mycophages have a partitioning

system: Redrock, DaVinci, Gladiator, Hammer and Blue7. It is not known whether these phage

homologues function in the same way as their bacterial counterparts, and characterizing the mechanism

of actions of these genes will give insight into the lysogenic state of mycophage DaVinci. Attempts at

knocking out parA have been unsuccessful, and in order to complement the parA knockout pNIT with

parA will be used. pNIT is an inducible plasmid allow control over levels of expression of the protein. A

parA-GFP fusion on pNIT will be constructed to perform real-time fluorescent microscopy experiments

to give insight into the function of parA.

Funding: HHMI

Katelyn Shaffer

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. John Rosenberg

FUNCTION WITHOUT STRUCTURE: STUDYING &#945;-SYNUCLEIN AS A MACROMOLECULAR

COSOLVENT (OSMOLYTE)

&#945;-synuclein is a natively unfolded protein whose deposits accumulate in several

neurodegenerative diseases, appropriately called “synucleinopathies;” e.g. Parkinson’s disease (PD).

&#945;-synuclein appears to be a pleitropic effector. One suggested role is in vesicular trafficking.

Another is based on the observation that it stimulates protein phosphatase 2A (PP2A) activity, both in

vivo and in vitro. Conventional thinking on natively unfolded proteins argues they adopt a folded state

when bound to a suitable macromolecular partner, preserving the “structure determines function”

paradigm. A crystal structure recently obtained in our laboratory and resulting work suggests a different

paradigm, one in which &#945;-synuclein can have a biochemical function while in a structurally

disordered state by acting as a macromolecular cosolvent (or osmolyte), where it participates in the

solvation of other proteins. We have been using dynamic light scattering (DLS) to characterize an

interaction between molecules of a GST- &#945;-synuclein fusion protein that appears to be mediated

by &#945;-synuclein. This biologically active fusion protein forms a (polydisperse) collection of particles

dominated by tetramers. The 2D HSQC NMR spectrum of the fusion protein shows that &#945;-

synuclein is unstructured under the conditions that support the formation of tetramers. Conventional

osmolytes perturb the size and behavior of the fusion protein in DLS. GST alone in DLS forms a well-

defined stable dimer. GST-fusions of &#945;-synuclein’s 3 modular domains show behavior unique to

each domain in DLS.Our results are consistent with the idea that disordered &#945;-synuclein

contributes to the formation of a composite solvation shell surrounding the GST.

Funding: HHMI

Aditee Shinde

Arts and Sciences

Faculty Mentor: Dr. Robert Gibbs

EFFECTS OF CHOLINERGIC LESIONS AND ESTRADIOL ON ESTROGEN RECEPTOR MRNA EXPRESSION IN

SPECIFIC REGIONS OF THE RAT BRAIN

Studies show that estrogen therapy can have beneficial effects on cognition in animals and humans, and

may protect against neurodegenerative disorders such as dementia and Alzheimer’s disease. However,

studies also suggest that beneficial effects of estrogen on cognition decrease with age and time

following menopause. Some of these diseases have been associated with signaling deficits in cholinergic

functions in the brain, yet estradiol has shown to significantly enhance cholinergic functions and

improve performance in many cognitive tasks. However, the mechanism through which estradiol can

enhance the cholinergic functions is unknown; perhaps it may exert its effects by binding to any of three

particular estrogen receptors, ERa (alpha), ERb (beta), and GPR30. For this project, I evaluated the

effects of basal forebrain cholinergic legions and estradiol treatments on levels of estrogen receptor

mRNA (messenger Ribosomal Nucleic Acid) in different regions of the brain. The hypothesis is that

cholinergic lesions lead to decreased estrogen receptor expression of mRNA in the brain, accounting for

the lack of estrogen.

Jessica Soe

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. William Saunders

IDENTIFICATION OF THE REQUIREMENT FOR MITOTIC MOTORS IN STRESS GRANULE DYNAMICS

Stress Granules (SGs) are cytoplasmic domains containing RNA binding proteins, 40S ribosome subunits,

stalled initiation complexes, and RNA aggregation that form under stressful conditions. The non-

translating messenger RNAs in SGs are being repaired, degraded, or stored due to initiation-related

damage. In order to identify how stress protein complexes form into SGs, we observed if microtubules

contribute to the highly dynamic nature of SGs. Microtubules have been shown to be required for SG

formation, which was confirmed in cells treated with sodium arsenate, which induces cells to form SGs

and there was a 30% decrease in SG assembly. Microtubule inhibitors were also tested for SG

dissolution. Cells treated with nocodazole, which disrupts microtubule polymerization, demonstrated

faster dissolution of SGs suggesting microtubules delay SGs disassembly. We investigated if mitotic

motors may function in SGs formation. Both mitotic motors, Eg5 and Kid were found to localize to SGs.

SiRNA studies were conducted and a 17% reduction in SG assembly was observed when Kid was knocked

down, while a 32% reduction was observed when Eg5 was knocked down, but Eg5’s ATPase activity was

not required for assembly. We inquired whether Eg5 helps dissociate SGs by treating cells with

monastrol, a specific inhibitor to Eg5, and a significant lag was observed during SG dissolution after Eg5’s

ATPase was inhibited. This demonstrates Eg5 is also required for SG dissolution and its ATPase activity is

required for this process. In summary, the identity of two mitotic motors in SG formation, coalescence

and or dissolution is a novel SG component and paves the way the identification of other families of

molecular motors in SG dynamics.

Funding: NIH

Yeohan Song

Neuroscience

Arts and Sciences

Faculty Mentor: Donald B. DeFranco, PhD

EFFECTS OF HYPERGLYCEMIA AND DEXAMETHASONE IN A MODEL OF NEURODEVELOPMENT

Background Hyperglycemia, or elevated blood glucose levels, in pregnant women can lead to

impairments in the development of fetal organ systems, including the nervous system. In diabetic

individuals, the hypothalamic-pituitary adrenal axis is overactive, leading to hypersecretion of

endogenous glucocorticoids. Glucocorticoid therapies have been shown to induce hyperglycemia in both

diabetic and non-diabetic patients, and elevated glucocorticoid levels have been found to mediate

deficits in cognitive function due to chronic stress. The exact mechanisms by which hyperglycemia and

glucocorticoids interact to affect neurodevelopment have yet to be determined. Methods Cell cultures

of the neuroblastoma line SH-SY5Y were used as an in vitro model to determine the impact of

hyperglycemia and glucocorticoids on neuronal growth and function. Results Undifferentiated SH-SY5Y

cells treated with &#8805;50 mM glucose exhibited lower viability after 24 h of treatment and showed

decreases in both Ki67 labeling and BrdU incorporation. Treatments of these cells with &#8805;100 nM

dexamethasone, a potent synthetic glucocorticoid, were associated with reductions in combined Ki67

labeling and BrdU incorporation. Future Directions The effects of hyperglycemia and dexamethasone on

apoptotic mechanisms and signaling pathways in SH-SY5Y will need to be further studied.

Funding: NIH Grant R01 DK078394

Craig Stevenson

Chemical and Petroleum Engineering

Swanson School of Engineering

Faculty Mentor: Prof. Goetz Veser

A NANOSCALE APPROACH TO CARBON CAPTURE

Anthropogenic CO2 emissions are widely recognized as a leading cause for climate change. Capturing

CO2 from concentrated point emissions, such as fossil fuel power plants, is hence crucial for curbing

atmospheric CO2 concentrations. However, current CO2 capture technologies, typically based on the

use of liquid amines, result in large energy penalties and strongly increased cost of energy (up to 85%).

In order to overcome some of the shortcomings of liquid amines, much work has recently focused on

the use of silica-based solid sorbent materials, either via impregnation with amines or via grafting of

amine groups onto silica. In particular nanostructured silica materials are of great interest due to very

high CO2 capture capacities of amine-impregnated "nanosilicas", in some cases even exceeding that of

liquid amines. In our work, we are evaluating novel hollow silica "nanobubbles" recently synthesized in

our laboratory for the confinement of CO2 sorbent materials. First results indicate that these materials

hold great promise for CO2 capture via nanoconfined liquid sorbents, with current sorption capacities of

up to 125 mg CO2/g sorbent and the potential for further increases based on tailoring of the silica

nanostructure. Materials synthesis, characterization, and CO2 sorbtion results will be presented in the

poster.

Funding: NETL

Kimberly Stuchul

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Michael Palladino

MODELLING TRIOSEPHOSPHATE ISOMERASE DEFICIENCY IN DROSOPHILA

Triosephosphate isomerase (TPI) deficiency is a progressive neurodegenerative condition most often

resulting in death before 5 years of age. This autosomal recessive disease is caused by missense

mutations and remains poorly understood. Previously it has been shown that the Drosophila TPI

missense mutation M80T (TPIsgk) causes protein instability leading to higher protein turnover. In order

to further investigate mutant TPI protein stability and understand the neurodegenerative aspect of TPI

deficiency, five TPI missense mutations known to cause human disease (I170V, E104D, C41Y, F240L, and

V231M) and a catalytically inactive mutation (K12M) were generated in Drosophila melanogaster using

genomic engineering. Briefly, the TPI locus was replaced with an attp and LoxP site using homologous

recombination gene targeting. PGE-TPI-attB plasmids bearing the wild type Drosophila TPI gene (control)

or various mutations were generated using site-directed mutagenesis. Additionally, we have engineered

PGE-TPI-attB capable of expressing the human TPI protein. Genomic engineering was performed by

using site-directed recombination via the attP-attB recombination system. The engineered loci will

allow in vivo analysis of each disease-causing mutation, which will lead to a better understanding of

human TPI deficiency pathogenesis.

Funding: NIH

Jiaxiang Tao

Civil and Environmental Engineering

Swanson School of Engineering

Faculty Mentor: Dr. Albert C. To

Computational Modeling of Nanoporous Aluminum

Nanoporous metals have gained a significant momentum for their substantial applications in today’s

new construction material because of their very low relative densities. However, previous research has

shown that nanoporous metals are weak, brittle, and undergoes a catastrophic failure when they are

subjected to tension. In order to prevent such “catastrophic failure”, Molecular Dynamics simulation is

applied to further investigate the failure mechanism of nanoporous aluminum, with different porosities,

under uniaxial tensile loading. The general computational results demonstrate that ligament size and

joint stiffness of nanoporous aluminum structure govern its softening rate. To be more specific, high

porosity structure with small ligaments and joints soften much slower and local stress is uniformly

distributed across the structure with no specific high stress localization and, thus, undergoes less

catastrophic failure. In contrast, for the low porosity structure with large ligaments and joints, stress is

highly localized on a small region, causing progressive softening and more catastrophic failure. More

evidences have proven that the joint size mainly governs the softening rates, because the high

compliance of joints allows the overloading stress, due to the rupture of weakest ligament, more

effectively spreads through the entire structure.

Funding: University of Pittsburgh Undergraduate Summer Research Internship Program

Hendrik van Hemmen

Civil and Environmental Engineering

Swanson School of Engineering

Faculty Mentor: Dr. Xu Liang

Design of Wireless Sap-flow Sensor Housings

Describes the design of a system to attach remote sap-flow sensor systems to trees at the Audubon

Center north of Pittsburgh. The system used prior to this design suffered from problems with water-

tightness, rigidity, animal damage, and bulkiness.

Funding: Swanson School of Engineering, REU program

Michael Volkwein

Civil and Environmental Engineering

Swanson School of Engineering

Faculty Mentor: Dr. Jorge D. Abad

COMPARISON AND ANALYSIS OF HYDRODYNAMIC MODELS FOR RESTORATION PROJECTS: THE CASE

OF POOL-RIFFLE STRUCTURES

Naturalization of urbanized streams through the use of pool-riffle structures creates a more hospitable

environment for fish and a consistent depth during high discharges while providing self-maintenance of

the structure. Whereas during low flow sediment builds up in pools, a pool-riffle structure with

sinusoidal narrowing banks allows higher shear stresses to develop in pools during high flows thereby

maintaining a pool-riffle structure over time. To design these self maintaining structures, hydrodynamic

models are used to optimize the parameters necessary to facilitate this shear stress reversing process.

1D models are often favored in place of resource intensive 3D models. However, the 1D and 3D pool-

riffle models do not always agree. 2-D models can be used to attempt to validate the 1D models and

describe their limitations. 1-D modeling was completed for four channel designs at eight levels of

discharge using HEC-RAS. The pool-riffle design displayed the self-maintenance mechanism for four flow

rates. Shear stresses in the pools did appear to exceed the stress at the riffles during high discharges. 2D

depth-averaged modeling was done using River2D and initial results show that the mechanism may not

be effective. This discrepancy should be compared to 3D models in more detail. Industry uses 1D

modeling most frequently for its cost effectiveness, showing that 1D models may not be sufficient in

pool-riffle design could be significant.

Funding: Swanson School of Engineering and the Heinz Endowment Award

Katelyn Walzer

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Jon P. Boyle

THE CODING REGION OF ROP43 MAY CONTRIBUTE TO INCREASED TRANSCRIPTION IN TOXOPLASMA

GONDII

Toxoplasma gondii infects over 60 million Americans and causes serious diseases in

immunocompromised hosts. These individuals suffer from the pathogenic effects of T. gondii when

chronic infections (caused by bradyzoites) are reactivated in T. gondii tissue cysts, initiating acute

infections (caused by tachyzoites). The ROP42 subfamily of genes is upregulated during tachyzoite-to-

bradyzoite differentiation and may therefore play a role in this process. Our goal is to determine the

minimal nucleotide sequence required for upregulation of these genes during differentiation by using

luciferase assays. We observed that including the first 270 nucleotides of the coding region of ROP42

increases the luciferase activity 21-fold during the conversion of tachyzoites to bradyzoites compared to

4.2-fold when using the promoter alone. This suggests that the nucleotide sequence in the 270 bp

region is involved in transcriptional regulation of ROP42 and/or that the peptide sequence it encodes

may alter the stability of the luciferase fusion protein. We examined whether a similar construct from a

closely related paralog, ROP43, behaved similarly in strain CTG of T. gondii. Our results indicate that

there is a 14.2-fold increase in the upregulation of luciferase activity when the promoter and 270 bp

fragment is present. This is in contrast to a 7.7-fold increase when only the promoter is present. Further

experiments will be done to determine if the upregulation caused by the 270 bp fragment works equally

well in a more virulent strain of T. gondii, ME49. We also want to find the minimal sequence within the

270 base pairs that causes an increase in transcript abundance, transcript stability, and/or protein

stability.

Funding: Summer Undergraduate Research Fellowship (HHMI-funded), Chancellor's Undergraduate

Research Fellowship (University Honors College)

David Wang

Chemistry

Arts and Sciences

Faculty Mentor: Dr. Lillian Chong

COMPUTER SIMULATIONS OF AN ARTIFICIAL ZYMOGEN DESIGNED TO COMBAT HIV

Molecular switches are biomolecules with functions that can be turned “on” or “off” in response to

signals from the surrounding environment such as pH, ligand binding, or enzymatic modification. One

strategy for engineering molecular switches involves the design of “alternate frame folding.” In this

design, two proteins are fused together such that they share an overlapping sequence, resulting in

alternate frames of the protein sequence that fold in a mutually exclusive fashion. This design has been

used to create a zymogen for the treatment of AIDS where the alternate frames correspond to two

different forms of barnase: one active and the other inactive. The inactive form can be shifted to the

active form upon cleavage by HIV protease, thereby unleashing the toxic enzymatic activity of barnase

only in HIV-infected cells. Since the “on” and “off” states of this switch are partially unfolded, it is

difficult to determine the structures of these states using X-ray crystallography or NMR spectroscopy. An

alternative approach is to use computer simulations with residue-level detail. Here we use such

simulations to characterize the “on/off” states of the barnase zymogen as well as its mechanism of

conformational switching. Understanding this mechanism can aid in the design of future molecular

switches.

Funding: University of Pittsburgh Startup Funds

Kurt Weiberth

Biological Sciences

Arts and Sciences

Faculty Mentor: Christopher J. Guerriero

Determining the Role of Chaperones in Endoplasmic Reticulum-Associated Degradation

Proper protein folding is vital for a cell’s survival and when secretory proteins misfold, one mechanism

to eliminate them is known as endoplasmic reticulum-associated degradation (ERAD). The ERAD of some

proteins has been linked to diseases; therefore, better understanding the mechanisms of ERAD can lead

to treatments of these diseases. In this study, the chaperones and components for ERAD were examined

using the protein Ste6p*, a native yeast protein with 12 transmembrane units that is a well-

characterized ERAD substrate. Currently, the relationship between chaperone requirements for ERAD

and the number of transmembrane domains in a substrate are unknown. The goal of this study was to

determine the chaperones required for substrates containing different numbers of transmembrane

domains but the same degradation signal. To do this, a chimeric protein, called Chimera A, was

constructed containing two transmembrane domains and the misfolded domain of Ste6p*. The rate of

degradation was monitored in yeast treated with a chemical inhibitor of ERAD. Following treatment,

Chimera A was stabilized by approximately 5-fold. This data supports the use of Chimera A to study the

specific chaperones and other requirements for ERAD.

Funding: Howard Hughes Medical Institute Summer Undergraduate Research Fellowship

Paul Werntges

Civil and Environmental Engineering

Swanson School of Engineering

Faculty Mentor: Dr. Piervincenzo Rizzo

OPTIMAL ANGLE DETERMINATION FOR THE DETECTION OF SYMMETRIC GUIDED WAVE MODES

Underwater structures that are used in everyday life include ships, submarines, and underwater pipes.

Very often the inspection of these structures requires temporary termination of service or the

employment of rugged technology that can withstand underwater conditions. In recent years the use of

Guided Ultrasonic Waves (GUWs) for the Nondestructive Evaluation (NDE) and the Structural Health

Monitoring (SHM) of waveguides has drawn much attention. In this study GUWs are exploited to

inspect plates immersed in water. The novelty of the present project is twofold: for the first time waves

are generated by means of pulsed laser; and the optimal angle of the sensing system for the detection

of the symmetric (So) mode is determined. A pulsed laser was used to generate stress waves in an

aluminum plate immersed in water, and an immersion piezoelectric transducer detected the

propagating waves. Signals were detected starting at 0° with respect to the vertical axis, and rotated

toward the point-source in three degree increments, up to 45°. The time waveforms were extracted

from an oscilloscope, and processed using Matlab software. Our results confirm theoretical predictions

of wave speed and dispersion frequencies for the symmetric S0 mode. Furthermore, our experimental

optimal angle conclusion is consistent with the prediction made using Snell’s Law. It is because of these

parallels that we can conclude that our experiment was a success.

Funding: NSF

Whitney White

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Howard Chang, MD,PhD, Stanford University

METASTASIS ASSOCIATED LUNG ADENOCARCINOMA TRANSCRIPT 1 ASSOCIATES WITH THE MLL

COMPLEX

While much has been discovered about RNA, long intergenic noncoding RNAs (lincRNAs) are still an area

of research that is vastly unexplored. In the past couple of years, thousands of lincRNAs have been

identified, yet the function of many of them still remain unknown. Of the long intergenic noncoding

RNAs that have been characterized, most serve a role in gene or epigenetic regulation. Malat-1 is a long

intergenic noncoding RNA that has been shown to be linked with increased metastasis in various

carcinomas. While it has potential to serve as a marker for survival, its function currently remains

unknown. Previous research suggested that Malat-1 might be associated with the MLL complex, a

protein complex that regulates epigenetic activation. In our studies, we have shown that Malat-1

associates with various proteins of the MLL complex using RNA immunoprecipitation. By identifying

Malat-1’s binding partners we hope to uncover some information about its function and the roles long

intergenic noncoding RNAs play in cancer.

Funding: HHMI ExROP

Emily Wolff

Civil and Environmental Engineering

Swanson School of Engineering

Faculty Mentor: Dr. Radisav Vidic

MARCELLUS SHALE FLOWBACK WATER TREATMENT FEASIBILITY STUDY WITH ACID MINE DRAINAGE

In recent years natural gas extraction from the Marcellus Shale formation, which underlies much of

Pennsylvania and some neighbouring states, has become feasible due to developments of the hydraulic

fracturing process. Hydraulic fracturing, known as “fracing” in industry, is an extremely water intensive

process requiring 3 to 5 million gallons of freshwater to frac one well on average. The reuse of water

recovered after the hydrofracturing, i.e. flowback, is limited by the low recovery, the high salinity and

expensive treatment costs. The reuse of flowback water requires the removal of toxic heavy metals

(barium, strontium) and other ions that can cause scaling and corrosion in the gas well. The objective of

this study is to develop a sustainable management approach for Marcellus flowback water that will

allow its reuse for hydrofracturing subsequent wells. In particular, the study is focusing on the feasibility

of mixing flowback water with acid mine drainage (AMD) which will serve as make-up water and remove

barium and strontium through sulfate precipitation. The work presented in this poster shows the initial

steps in understanding the chemistry of different flowback waters and AMD mixes by studying

precipitation kinetics and crystal size, morphology and composition.

Funding: U.S. Department of Energy

Narayana Yelleswarapu

Psychology

Arts and Sciences

Faculty Mentor: Dr. Anthony E Kline

The effects of the 5-HT1A and 5-HT7 receptors on behavioral recovery after traumatic brain injury

Previous data from our laboratory has shown that the 5-HT1A receptor agonist 8-OH-DPAT improves

behavioral recovery after experimental traumatic brain injury. It has generally been thought that 8-OH-

DPAT is mediating its beneficial effects via the 5-HT1A receptor. However, because 8-OH-DPAT also has

partial agonist activity at the 5-HT7 receptor, it is possible that activation of this receptor may be playing

as role in the benefits observed. To elucidate this possibility, we evaluated the effects of antagonists for

the 5-HT1A and 5-HT7 receptors. Specifically, sixty-eight rats received either a controlled cortical impact

(CCI) or sham injury and then were randomly assigned to the following groups: 8-OH-DPAT (0.5 mg/kg),

8-OH-DPAT + SB 269970 HCl (5-HT7 antagonist; 2 mg/kg), 8-OH-DPAT + WAY 100635 (5-HT1A

antagonist; 0.5 mg/kg), or saline vehicle (1 mL/kg). Each group was tested on motor (beam) and

cognitive (Morris water maze) tasks on days 1-5 and 14-19, respectively. 8-OH-DPAT alone, or when

combined with the antagonists showed cognitive improvement relative to vehicle controls, but there

was no statistical difference among the groups. The data did not provide the clear cut distinction of

what receptor is mediating the benefits observed with 8-OH-DPAT. It is possible that both receptors are

playing a role. Evaluation of a 5-HT7 receptor agonist is warranted to further elucidate the potential

mechanisms and our understanding of 8-OH-DPAT and its effects.

Funding: NIH

Ally Young

Biological Sciences

Arts and Sciences

Faculty Mentor: Dr. Karen Arndt

INVESTIGATING THE FUNCTION OF RKR1 THROUGH A HISTONE-BASED GENETIC SCREEN IN S.

CEREVISIAE

Protein ubiquitylation regulates many important processes in eukaryotes. Uncontrolled ubiquityation

has been associated with a number of diseases, including cancer. Rkr1 is a previously unstudied

ubiquitin ligase that has been linked to transcription and chromatin function in yeast. To elucidate a

function for Rkr1, we devised a screen to find genetic interactions between Rkr1 and histone mutations

using a plasmid library encoding alanine substitutions at all histone residues. Each of these plasmids was

transformed into a rkr1 deletion strain, which also contained a plasmid encoding the only wildtype

source of histone in the cell. Transformants were forced to lose the wildtype copy by counter-selection.

We hypothesize that a histone mutation that affects a process for which Rkr1 also plays a role will result

in a genetic interaction when both Rkr1 and the histone residue are compromised. The screen has

recently been completed for histones H2A and H2B. Candidates found were re-transformed, subjected

to dilution assays, and sequenced to verify mutations. Of the 12 candidates identified, 2 are known

lethal mutations. Another is the mutation of H2B K123, which is known to be synthetically lethal with

the deletion of rkr1. Interestingly, many candidates have published phenotypes associated with defects

in transcription, including sensitivities to hydroxyurea, 6-azauracil, and mycophenolic acid. The

candidate residues also form an obvious patch on the surface of the histone molecule when mapped.

Future analysis will investigate the role of Rkr1 in pathways associated with these mutations. Finally, the

entire screen will be repeated for histones H3 and H4.

Funding: Arts and Sciences Undergraduate Research Award